APPLICATION OF NITRIC OXIDE RESEARCH TO DRUG DEVELOPMENT

Abstract

The role of nitric oxide in cellular signaling in the past three decades has become one of the most
rapidly growing areas in biology. Nitric oxide (NO) is a gas and a free radical with an unshared
electron that can regulate an ever-growing list of biological processes. Nitric oxide is formed from
L-arginine by a family of enzymes called nitric oxide synthases. These enzymes have a complex
requirement for a number of co-factors and regulators including NADPH, tetrahydrobioterin, flavins,
calmodulin and heme. The enzymes are present in most cells and tissues. In many instances, nitric oxide mediates its
biological effects by activating the soluble isoform of guanylyl cyclase (SGC) and increasing cyclic GMP synthesis from
GTP Cyclic GMP, in tum, can activate cyclic GMP-dependent protein kinase (PKG) and can cause smooth muscles and
blood vessels to relax, decrease platelet aggregation, alter neuron function, etc. These effects can decrease blood
pressure, increase blood flow to tissues, alter' memory and behavior, decrease blood clotting, etc. The list of effects of
nitric oxide that are independent of cyclic GMP formation is also growing at a rapid rate. For example, nitric oxide can
interact with transition metals such as iron, thiol groups, other free radicals, oxygen, superoxide anion, unsaturated fatty
acids, and other molecules. Some of these reactions result in the oxidation of nitric oxide to nitrite and nitrate to
terminate the effect and perhaps act as NO reservoir for future NO formations; while other reactions can lead to altered
protein structure function and/or catalytic capacity. These effects of (NO) probably regulate bacterial infections,
inflammation of tissues, tumor growth, and other disorders. These diverse effects of nitric oxide that are cyclic GMP
dependent or independent can alter and regulate numerous important physiological events in cell regulation and
function. Nitric oxide can function as an intracellular messenger, an autacoid, a paracrine substance, a neurotransmitter,
or as a hormone that can be carried to distant sites for effects. Thus, it is a unique molecule with an array of signaling
functions. However, with any messenger molecule, there can be too little or too much of the substance, resulting in
pathological events. Some of the methods to regulate either nitric oxide formation, metabolism, or function have been in
clinical use for more than a century as with the use of organic nitrates and nitroglycerin in angina pectoris that was
initiated in the 1870’s. Inhalation of low concentrations of nitric oxide can be beneficial in premature infants with
pulmonary hyperension and increase survival rates. Ongoing clinical trials with nitric oxide synthase inhibitors and nitric
oxide scavengers are examining the effects of these agents in septic shock, hypotension with dialysis, inflammatory
disorders, cancer therapy, etc. Recognition of additional molecular targets in the areas of nitric oxide and cyclic GMP
research will continue to promote drug discovery and development programs in this field. Current and future research
will undoubtedly expand the clinician's therapeutic armamentarium to manage a number of important diseases by
perturbing nitric oxide formation and metabolism. Such promise and expectations have obviously fueled the interests in
nitric oxide research for a growing list of potential therapeutic applications. There have been and will continue to be
many opportunities from nitric oxide and cyclic GMP research to develop novel and important therapeutic agents. There
are presently more than 150,000 publications in the areas of nitric oxide research. The lecture will discuss our discovery
of the first biological effects of nitric oxide and how the field has evolved since our original reports in 1977. The
possible utility of this signaling pathway to facilitate novel drug development and the creation of numerous projects in
the Pharmaceutical and Biotechnology industries will also be discussed.

CONFERENCE ABSTRACT (PLENARY)

FROM STRUCTURES TO ADVANCED THERAPEUTICS, AND ABOUT ORIGIN OF LIFE

Abstract

Ribosomes, the universal cellular machines for translation of the genetic code into proteins, possess spectacular architecture accompanied by inherent mobility, allowing for their smooth performance as polymerases that translate the genetic code into proteins. The site for peptide bond formation is located within a universal internal semi-symmetrical region. The high conservation of this region implies its existence irrespective of environmental conditions and indicates that it may represent an ancient RNA machine. Hence, it could be the kernel around which life originated. The mechanistic and genetic applications of this finding will be discussed

Owing to the key role played by ribosomes in life cycles, almost half of the clinically useful antibiotics paralyze ribosomes by binding to their functional sites. By investigating the three dimensional structures of ribosomes from nonpathogenic bacteria as models for genuine pathogens, common features were identified. Thus, the antibiotics binding modes, inhibitory actions and synergism pathways have been determined for almost all ribosomal antibiotics. These indicated the principles of differentiation between patients and pathogens and suggested common principles of mechanisms leading to bacterial resistance.

The incredible global increase in resistance to antibiotics that we are witnessing recently is a serious medical threat. It seems that the world is approaching a post-antibiotic era, in which common infections and minor injuries that have been treatable for decades could become fatal once again.

As species specific diversity was detected in susceptibility to infectious diseases and in developing specific resistance mechanisms, our structural studies have been extended to ribosomes from genuine pathogens. By determining the high resolution structure of the first and only ribosomal particle from a genuine pathogen with several antibiotics, we identified subtle, albeit highly significant structural elements that can account for the species specificity in resistance, thus could paved ways for improvement of existing antibiotics as well as for the design of advanced therapeutics capable of minimizing antibiotics resistance.

CONFERENCE ABSTRACT (PLENARY)

MicroRNAs, SMALL INTERFERING RNAs AND MODULAR THERAPEUTICS

Abstract

RNA interference was discovered a little over ten years ago and subsequently was shown to be mediated by intracellular
double stranded small RNAs approximately 21 nucleotides in length (siRNA). In mammals, these RNAs enter the
microRNA pathway, present in all cell types, are loaded into a complex containing the critical Argonaute protein, and
target its activity to cleave and cause degradation of specific complimentary mRNA. Thus, in principle with the
appropriate design of the siRNA any target gene could be silenced. Over the past years, the challenge of effective
delivery of the hydrophilic siRNAs to cells has been advanced to where it is now possible in a therapeutically attractive
fashion to silence genes expressed in the liver of humans with a sugar-based conjugate of a chemically modified siRNA.
These therapeutic agents are modular in structure, where one constituent provides a gene-specific component whose
modulation of expression is beneficial, while the other agent targets and facilitates delivery to the inside of cells. This
modular property greatly reduces the time required to develop therapeutics to new disease modifying genes and is an
example of future developments in pharmaceuticals. Examples of these types of agents will be discussed. The activities
of small RNA based agents will be set in the context of the known biology of small non-coding RNAs.

CONFERENCE ABSTRACT (PLENARY)

EVOLUTIONARY DYNAMICS AND TREATMENT OF CANCER

Abstract

Cancer is an evolutionary process. Cancer initiation and progression are caused by somatic mutation and selection of dividing cells. The mathematical theory of evolution can therefore provide quantitative insights into human cancer. I will discuss the role of chromosomal instability (CIN) and the accumulation of drivers and passengers in growing tumors. I will study success and failure of targeted therapy including combination of different drugs and evolution of resistance. A simple conclusion is that combination treatment can succeed, if the cancer requires at least two point mutations to gain resistance. From the perspective of preventing resistance, simultaneous therapy is highly recommended whereas sequential therapy is a recipe for almost certain treatment failure.

Abstract

Endogenous mechanisms controlling inflammation are of paramount importance because persistent and chronic inflammation can impact all organs and tissues throughout the body and are involved in many widely occurring diseases. Recent advances in our appreciation of the molecular mechanisms in resolution of acute inflammation (RoI) and ischemia-reperfusion injury systematically uncovered a novel genus of potent pro-resolving autacoid families, each biosynthesized from essential polyunsaturated fatty acids (PUFA) to activate potent responses not shared by the substrate. These include the resolvins (Rv), protectins (PD) and maresins (MaR), collectively termedspecialized proresolving mediators (SPM) that act in pico-nanogram range. SPM are temporally and spatially biosynthesized by resolving-inflammatory exudates, which proved to evoke potent antiinflammatory and pro-resolving actions as well as enhance microbial clearance. The potent SPM actions and complete structures are confirmed, which also permitted use of LC-MS-MS-based metabololipidomics to identify SPM in human and murine tissues (i.e. peripheral blood, breast milk, adipose, lymphoid, placenta), isolated human cells types (e.g. apoptotic human neutrophils, microparticles and macrophage phenotypes M1, M2), fish and diminished SPM in human pathologies e.g. breath condensates, Alzheimer brain, and synovial fluids from rheumatoid patients (CN Serhan Nature June vol 510, 2014 doi:10.1038/nature13479). Specific SPM demonstrate potent and stereoselective actions that involve specific G-protein-coupled receptors and are not immunosuppressive. Lipid mediator-metabololipidomics with selflimited resolving inflammatory exudates and human tissues demonstrated temporal orchestration of the SPM, i.e. RvD1 and RvD2 antecede RvD3, and MaR1 in mice and human tissues (Colas et al. AJP 2014). Many of the SPM born in inflammation-resolution are now shown to have potent actions and roles within host defense against bacteria and virus, pain, organ protection, tissue regeneration, exercise and neurobiology/cognitive function. This Plenary Lecture will update advances in SPM mechanisms in RoI, their new sites of formation and novel actions that opened the door for their role(s) in resolution physiology and pharmacology. Together, these new SPM families provide opportunities for resolution-based pharmacology and resolution physiology.

CONFERENCE ABSTRACT (PLENARY)

Designing
Biology
for
a
Healthy
World

Pamela
A.
Silver
Department
of
Systems
Biology,
Harvard
Medical
School
and
The
Wyss
Institute
for
Biologically
Inspired
Engineering,
Harvard
University

Abstract

The engineering of Biology presents infinite opportunities for therapeutic design, diagnosis, and prevention of disease. Towards these goals, we seek to make the engineering of Biology faster, more predictable and cheaper. This ‘Synthetic Biology’ has deep practical and social consequences for the pharmaceutical as well as the commodity industry. Here, I will present concepts and experiments that begin to address how we approach these problems in a systematic way.

By one strategy, we seek to predictably engineer mammalian cells to produce novel compounds that could potentially act as new therapeutics. For example, we have developed an algorithm for biosynthesis of new steroids that could have increased specificity towards their respective targets. This has implications in treatment of inflammation and clean production of other chemicals of interest.

By a second strategy, we design chimeric proteins to act as specific therapeutics. Specificity in biologics remains one of the outstanding issues in their use. We have again developed an algorithm based on coarse grain modeling for the predictable design on new proteins. Some have been tested in animals and show the predicted effects.

Lastly, we engineer components of the microbiome to act as both diagnostics and therapeutics. In one example, we have engineered natural gut bacteria to record the exposure of animals to antibiotics and to count the number of cell divisions as the bacteria passes through the gut. We can engineer the same bacteria to secrete toxins that could result in localized killing of pathogens and to act in a communal manner. Taken together, these experiments have far-reaching implications for the use of biology to prevent and treat disease in the future.

CONFERENCE ABSTRACT (PLENARY)

RECONSTRUCTION, MODELING AND USE OF GENOME-SCALE NETWORKS IN BIOLOGY

Bernhard Palsson

Departments of Bioengineering and Pediatrics University of California, San Diego, San Diego, USA

Abstract

Following the availability of full genome sequences in the mid 1990s, an effort was initiated to
reconstruct, on a genome-scale, the biochemical reaction networks that underlie cellular functions.
After 15 years of intense efforts, we now have highly curated network reconstructions, their
experimental validation, and the generation of mathematical and modeling procedures available that
allow the computation of cellular functions from genome- and bibliome-wide data sets. This effort
has put a mechanistic basis into the most fundamental relationship in the life sciences; the genotypephenotype
relationship. This effort has started with simple organisms and the best characterized cellular functions and it
is steadily growing in scope and biological complexity.

VBP15 FOR DMD: A COLLABORATIVE PUBLIC/PRIVATE PARTNERSHIP ENABLED BY VENTURE PHILANTHROPY SUPPORT FROM THE INTERNATIONAL STAKE-HOLDER COMMUNITY

Abstract

Drug development is a lengthy and time consuming process, where it has been suggested that it typically costs $500 million and takes 15 years to bring a drug to market. The high costs become problematic with orphan drugs, where there may be very few patients to prescribe the new drug and distribute costs. Also, there are increasing opportunities for highly targeted drugs, but often in ever more stratified patient populations, so the proportion of orphan drugs is expected to increase dramatically. Methods to decrease the costs and time associated with orphan drug development include early de-risking (reducing late stage expensive failures), and accelerated approval (moving larger efficacy trials to the post-marketing space). VBP15 is a first in human drug developed for Duchenne muscular dystrophy (DMD). Intellectual property for the program was transferred from Children’s National Medical Center to ReveraGen Biopharma. Initial seed funding for lead compound selection was accomplished through support of the Department of Defense CDMRP, and early de-risking of the lead (VBP15) was done in partnership with the National Institutes of Health TRND program. Pre-clinical studies and Phase 1 trials were funded by venture philanthropy support of seven DMD foundations through a risk sharing and profit sharing model based on later drug sales. Phase 2a and 2b trials in DMD patients are to run by international academic networks, with the possibility of accelerated approval after a short-term (3-6 month) trial with a measure of strength as the primary outcome measure. VBP15 is a steroidal compound built on a delta 9,11 backbone, where the drug shows a high affinity for the glucocorticoid receptor, but is effective in dissociating anti-inflammatory transrepression subactivities (retention of efficacy), from the transactivation subproperties associated with side effects (growth stunting, adrenal suppression, immune suppression). VBP15 has shown efficacy in pre-clinical models of multiple chronic inflammatory states, including allergic lung disease, arthritis, multiple sclerosis, inflammatory bowel disease, and sickle cell anemia. Thus, VBP15 holds potential for replacing traditional glucocorticoids in multiple indications, including DMD.

Abstract

The discovery of new antibiotics has become urgent as a result of the emergence of resistance and new pathogenic bacterial strains. However, this need has coincided with unprecedented lowering of levels of productivity in the drug discovery process and consequent reduced investment from large pharma. New strategies for antibacterial drug discovery are required, and a renewed understanding of the value of a natural products’ guided approach has emerged. Our focus has been on the use of antibacterial natural products containing tetramate core structures, and using equisetin and reutericyclin as inspiration, we have developed novel chemistry that uses suitable serine, threonine and cysteine-derived oxazolidine templates for highly chemo- and diasteroselective ring closure reactions leading to tetramic acid derivatives. Although simple unsubstituted pyrrolidines and tetramates appear to be intrinsically devoid of activity, application of these templates for fragment-based synthesis, has permitted access to several compound series which possess high levels of antibacterial activity, SAR analysis has permitted some optimization of the initial activity and MOA and other pharmacokinetic data been obtained.

This lecture will illustrate the potential of natural products to guide antibacterial drug discovery, the role of synthetic organic chemistry in the construction of libraries which mimic these natural products, and suggest a possible way forward for more efficient drug discovery strategies.

ENGINEERED HYDROGELS FOR REGENERATIVE MEDICINE APPLICATIONS

Abstract

Engineered materials that integrate advances in polymer chemistry, nanotechnology, and biological sciences have the potential to create powerful medical therapies. Our group aims to engineer tissue regenerative therapies using water-containing polymer networks, called hydrogels, that can regulate cell behavior. Specifically, we have developed photocrosslinkable hybrid hydrogels that combine natural biomolecules with nanoparticles to regulate the chemical, biological, mechanical and electrical properties of gels. These functional scaffolds induce the differentiation of stem cells to desired cell types and direct the formation of vascularized heart or bone tissues. Since tissue function is highly dependent on architecture, we have also used microfabrication methods, such as microfluidics, photolithography, bioprinting, and molding, to regulate the architecture of these materials. We have employed these strategies to generate miniaturized tissues. To create tissue complexity, we have also developed directed assembly techniques to compile small tissue modules into larger constructs. It is anticipated that such approaches will lead to the development of next-generation regenerative therapeutics and biomedical devices.

Abstract

Multidrug ABC (“ATP-binding cassette”) transporters are involved, upon overexpression, in chemoresistant tumors by pumping anticancer drugs out of the cells. For early discovered ABCB1/ “P-glycoprotein”, third-generation drug-efflux inhibitors are under clinical development. For more recently identified ABCG2/“breast cancer resistance protein”, we have screened different series of flavonoids and derivatives, such as flavones, rotenoids and acridones, and more recently chalcones [1, 2], chromones [3, 4], and indenoindoles [5], as inhibitors of mitoxantrone efflux from transfected HEK293 human cells and chemosensitizers of cell proliferation, to establish 3D-Quantitative Structure-Activity Relationships. Two types of selective, non-competitive, inhibitors have been characterized, either inhibiting or stimulating the basal ATPase activity. The most potent one is indeed efficient in vivo on SCID mice, xenografted with human ABCG2-transfected cells, by chemosensitizing tumor growth to the drug-substrate irinotecan [6]. These selective inhibitors constitute good drug candidates, with low intrinsic toxicity, as sensitizers of cell proliferation to conventional chemotherapeutics.

The “Multidrug Resistance Protein 1” ABCC1 is able to catalyze the efflux of either glutathione conjugates or free glutathione together with hydrophobic substrate drugs. We have identified modulators such as verapamil [7, 8] mimicking substrates and inducing a fast and massive efflux of intracellular glutathione from ABCC1-overexpressing cells, leading to a selective cell death through apoptosis, due to “collateral sensitivity”, or hypersensitivity. The overexpressed transporter then constitutes the Achilles’heel of such resistant cancer cells. Since verapamil is known for its cadiotoxic effects, we investigated other types of modulators such as xanthones, flavones [9] and flavonoid dimers. Glutathione efflux appeared to be necessary, but not sufficient alone, to trigger apoptosis, indicating the contribution of other partner(s) or signaling pathway(s). Such apoptosis inducers may constitute a new type of anticancer drugs operating through an original strategy aimed at selectively targeting and eliminating multidrug-resistant tumors overexpressing the ABCC1 transporter [10].

ON THE ORIGIN AND COMPLETENESS OF LIGAND BINDING POCKETS WITH
APPLICATIONS TO DRUG DISCOVERY

Abstract

The intrinsic ability of protein structures to exhibit the geometric and sequence properties required
for ligand binding without evolutionary selection is shown by the coincidence of the properties of
pockets in native, single domain proteins with those in computationally generated, compact
homopolypeptide, artificial structures, ART. The library of native pockets is covered by a
remarkably small number of representative pockets (~400), with virtually every native pocket having
a statistically significant match in the ART library, suggesting that the library is complete. When
sequences are selected for ART structures based on fold stability, pocket sequence conservation is coincident to native.
The fact that structurally and sequentially similar pockets occur across fold classes combined with the small number of
representative pockets in native proteins implies that promiscuous interactions are inherent to proteins. Based on
comparison of PDB and ART structures and pockets, the widespread assumption that the co-occurrence of global
structure, pocket similarity, and amino acid conservation demands an evolutionary relationship between proteins is
shown to significantly underestimate the random background probability. Indeed, many features of biochemical function
arise from the physical properties of proteins which evolution likely fine-tunes to achieve specificity. This study
suggests that a repertoire of thermodynamically (marginally) stable proteins could engage in many of the biochemical
reactions needed for living systems without selection for function, a conclusion with significant implications for the
origin of life. Finally, examples of experimental validation of promising small molecule hits that exploit the degeneracy
of ligand binding pockets are presented.

Enabling Technologies (Track)

NEW TECHNOLOGIES FOR IMPROVED VACCINES AGAINST INFECTIOUS DISEASES AND
CANCER

Abstract

Vaccines are without a doubt the most successful of mankind’s medical interventions. However,
despite more than two centuries of effective use of vaccines, many substantial challenges remain.
These include: 1) improvement of existing but suboptimal vaccines (e.g., tuberculosis, influenza), 2)
discovery and development of new vaccines against targets to address large unmet medical needs
(e.g., HIV, malaria, cancer), and 3) rapidly responding to new pathogens (e.g., newly emerging
microbes, bioweapons). Recent advancements have demonstrated proof of concept for active immunization in the
treatment of cancers. Taking full advantage will require the application of new technologies and paradigms in the areas
of tumor antigen identification and optimization, novel potent and safe adjuvants, and enhanced vaccine delivery
systems.

Drug Discovery in Preclinical Research (Track)

George Washington University, School of Medicine and Health Sciences, Washington D.C., USA

Abstract

Prior reports of hypomagnesemia in patients receiving the epidermal growth factor receptor (EGFR)-
blocking drug, Cetuximab, suggested that EGFR-tyrosine kinase inhibitors (TKI), like erlotinib
(Tarceva), may have a similar side effect. In addition, the possibility of preventing complications of
therapy by blockade of neurogenic inflammation was studied. Rats were treated with erlotinib for up
to 9 weeks and were investigated for hypomagnesemia, inflammation and cardiac dysfunction.
Plasma magnesium decreased progressively (-9 to -26%) between 3-9 weeks, and this was associated
with modest increases (+27% at 3 and + 25% at 9 weeks) in circulating levels of the neuropeptide, substance P (SP). In
previous work the TKI, tyrphostin AG-1478, caused similar effects. The superoxide-generating activity of circulating
neutrophils from erlotinib-treated rats increased 3-fold, and the plasma oxidative marker, 8-isoprostane rose 2.1-fold,
together with appearance of cardiac peri-vascular nitrotyrosine. The SP receptor blocker, aprepitant (Emend), attenuated
erlotinib-induced hypomagnesemia by 42% and completely prevented the rise in circulating SP; the increases in
neutrophil superoxide activity and 8-isoprostane were also suppressed. Echocardiography revealed mild to moderate
systolic dysfunction by 7 weeks of erlotinib treatment, with slight decreases in left ventricular ejection fraction (LVEF:
-11%) and % fractional shortening (%FS: -17%); the mitral valve E/A ratio was significantly reduced (-17.5%, week 9),
suggesting diastolic dysfunction. Also thinning of the left ventricular posterior wall (LVPWd & s in diastole and systole)
was detected by 7 - 9 weeks. Most interestingly, the co-administration of aprepitant attenuated all of the erlotinibinduced
effects on LV systolic, diastolic and anatomical parameters: At 9 weeks, LVEF improved 87.2%; LV % FS
improved 86.4%; and mitral valve E/A ratio improved 84.6%.

In conclusion, prolonged erlotinib treatment in rats induced moderate hypomagnesemia, along with SP-mediated
oxidative/inflammation stress, and mild to moderate cardiac dysfunction. These changes were substantially prevented by
SP receptor blockade implicating the novel role of neurogenic inflammation due to EGFR-TK inhibition.

CONFERENCE ABSTRACT (KEYNOTE)

Hot Topics in Natural Products (Track)

NATURE-BASED DRUG DISCOVERY – REEMERGING PARADIGM

M. Iqbal Choudhary and Atta-ur-Rahman

International Center for Chemical and Biological Sciences, (H.E.J. Research Institute of Chemistry
and Dr. Panjwani Center for Molecular Medicine and Drug Research) University of Karachi,
Karachi-75270, Pakistan

Abstract

Modern drug development is an expensive and lengthy process, which requires over $ 1.8-2.0 billion
worth of investment, and focused work of a large interdisciplinary team of scientists, involving years
of studies, and screening of a large chemical space. Unfortunately this situation has out-numbered
and out-resourced the academic institutions and pharmaceutical R & D of developing nations. The
role of academic institutions in drug development, particularly in developing countries, is gradually diminishing. As a
result, several diseases, affecting the lives of poor population of the South remain untreated. This situation demands a
major soul searching by pharmaceutical scientists who wish to serve the humanity through the skills they posses.
Overall change in paradigm in drug development is required, which create space for academic researchers and R & D
workers of developing nations to contribute in the discovery and development of drugs against diseases affecting their
regions. This change must involve the effective use of indigenous knowledge and resources. Natural products and their
traditional uses can play a very important role in drug development for poors by researchers of developing world. During
the presentation, results of our studies on natural products will be presented to support the argument that the knowledgebased
research on the natural products is a key to discover potential drug candidates at low cost.

Multidrug resistance is a challenging problem for the healthcare sector and is very common in familiar pathogens, such
as vancomycin-resistant enterococci and Staphylococcus aureus. Exposure and inappropriate use of the antibiotics is the
measure cause of MDR, both in developed and developing regions. Our study, focusing on the discovery of natural and
synthetic compounds, active against multidrug resistant bacteria Staphylococcus aureus and Pseudomonas aeruginosa have resulted in the identification of several novel and potent inhibitors of MDR Staphylococcus aureus, (EMRSA-17,
EMRSA-16, MRSA-252, and Pak clinical isolates) from natural sources. Resistance-reversal studies at molecular level
were carried out by employing flow cytometric and microscopic techniques. Synergistic and partial synergistic effects of
these compounds, in combination with antibiotics, were investigated. This work has so far resulted in the identification
of several novel “helper molecules”, which can increase the efficacy of existing antibiotics to over 1000-fold in some
cases.

Diabetes is one of the largest threats for public health in the new millennium, and its impact is felt most severely in
developing countries. It is a chronic disease that occurs when the pancreas does not produce enough insulin, or when the
body cannot effectively use the insulin it produces. Hyperglycaemia, or raised blood sugar, is a common feature of
uncontrolled diabetes which leads to serious damage to many of the body's systems, especially the nerves and blood
vessels. Diabetic patients are prone to long-term complications, such as retinopathy, cataract, atherosclerosis,
neuropathy, nephropathy and impaired wound healing. Most of these complications are related to standard changes in
tissues proteins by their non-enzymatic bindings with sugars molecules. Current treatments of diabetes are largely
ineffective in achieving the normal sugar levels and delaying the onset of late diabetic complication. Therefore, there is
an urgent need for new strategies for the treatment of diabetes, and its prevention against complications.

Among different therapeutic interventions, the discovery of effective α-glucosidase inhibitors and antiglycating agents
are considered to be the most important one, based on modern knowledge about the disease at molecular level. Primary
focus of these studies has been to discover lead molecules by using appropriate conventional and mechanism-based
biological screening techniques. As a result, a large number of potent antiglycation agents, and α-glucosidase inhibitors
of natural origin were discovered and structure-activity relationship studies were conducted. Many of these compounds
represent new examples of inhibitors of α-glucosidase and protein glycation.

Business Development (Track)

SESSION SPEAKER

CONFERENCE ABSTRACT (SESSION)

Business Development

FINANCIAL VALUATION ALGORITHM FOR THE ASSESSMENT OF THE FUTURES SALES
OF A NEW, INNOVATIVE MEDICINAL PRODUCT

Mark Nuijten

Health Care Valuation, Amsterdam, The Netherlands

Abstract

Registration is not the sole success factor anymore for future sales and valuation of the share of
company. Reimbursement procedures and the introduction of new business models like value-based
pricing and risk sharing agreements, have financial consequences for biotech companies. As the
future financial performance of a biotech company is directly related to the revenues of new products,
an appropriate assessment of the potential sales forecast of the portfolio of forthcoming new products
is an important predictor of the financial value of a pharmaceutical company. Therefore information on positive clinical
trials results of a new product should be followed by a reimbursement scan and a sales forecast model including the key
global markets.

The objective of this lecture is to present a financial valuation algorithm for the assessment of the futures sales of a new,
innovative medicinal product based on the current reimbursement policies and future business models for
reimbursement. The algorithm consists of a number pathways leading to an assessment of the market potential of the
new drug: no potential, limited potential, moderate potential and expected potential. The algorithm is applied to possible
new innovative products for treatment of depression and multiple sclerosis.

CONFERENCE ABSTRACT (SESSION)

Industrial and Manufacturing

Key Laboratory of Molecular Biophysics, The Ministry of Education, College of Life Science and
Technology, Huazhong University of Science and Technology, Wuhan 430074, P. R. China

Abstract

Aimed to major bottlenecks of high cost and low operation stability of free lipases, Candida rugosa
lipase (CRL) and Rhizopus oryzae lipase (ROL) were co-displayed on the cell surface of Pichia
pastoris and used as a whole-cell catalyst to produce biodiesel from tallow seed oil in this study.
After screened by double resistance and tributyrin medium, the resultant co-displayed recombinant GS115/pRCS with
the maximum activity of 470.59 U/g dry cells, being 3.9- and 1.3- fold compared with that of the single displayed ROL
and CRL1, respectively. The analysis of fluorescence microscope and Flow Cytometer demonstrated that ROL and
CRL1 were successfully co-displayed on the surface of recombinant P. pastoris GS115. When the self-immobilized
lipases were utilized as whole cell catalysts, the rate of methyl ester from co-displayed recombinants strain GS115/pRCS
harboring ROL and CRL1 is 1.4-fold compared with that of single displayed ROL. All these results indicate that
biodiesel catalyzed by the co-displayed enzymes with synergetic effect could be an alternative strategy for producing
biodiesel in low cost.

Industrial and Manufacturing (Track)

RHODOCOCCUS ERYTHROPOLIS NTU-1: FROM BIO-DEGRADATION TO BIOFLOCCULATION
AND BIO-DEMULSIFICATION

Abstract

An alkane-biodegrading bacterium identified as Rhodococcus erythropolis (NTU-1 strain) was
isolated from petroleum-contaminated soil. When long-chain alkanes are supplied as the carbon
source, NTU-1 tends to form pellets, ranging from 0.1 to 2 cm in diameter, and thus remove
significant amount of alkanes by biodegradation and physical trapping in a short period.
Quantitatively, more than 95% of each alkane (~2000ppmv) could be efficiently removed within 40–68 h. Further, Rhodococcus erythropolis strain NTU-1 was also confirmed to be able to degrade C10–C32 n-alkanes in diesel oil or
crude oil. While degrading these n-alkanes, NTU-1 also trapped most other non-degradable oil constitutes in pellets. In
batch cultures with 10,000ppmv diesel or crude oil, approximately 90% oil removal was achieved within 4 days. In
bioreactors with aeration and pH adjustment, an intermittent feed of 42,000ppmv n-hexadecane resulted in
approximately 87% removal within 4 weeks and an intermittent feed of 35,000ppmv diesel or crude oil resulted in more
than 90% removal within 2 weeks.

It was also found that, if NTU-1 was properly cultured and heat-dried, the cells could work as bio-flocculating agent that
could shorten the clean up time to 12h, as compared to 40-68h. Meanwhile, the cells further showed good capability to
de-emulsify oil/water/surfactant emulsion system and may play a role in separate oil from tertiary recovery in petroleum
industry.

These interesting and versatile phenomena of Rhodococcus erythropolis NTU-1 involving bio-degradation, bioflocculation
and bio-demulsification will be presented and discussed.

Industrial and Manufacturing (Track)

Centre for Research in Molecular Medicine, the University of Lahore, Lahore, Pakistan

Abstract

The study aims at establishing the biological assays for recombinant growth hormone (GH) of the
local Beetal breed of bovidae specie; caprine. For this purpose, a simple and highly sensitive
competitive enzyme immunoassay for recombinant caprine growth hormone (rcGH) was optimized.
Antiserum was raised in two local breed rabbits and horseradish peroxidase was labeled with rcGH.
96-well microtitre plates were coated with primary antibody (rabbit anti-rcGH). The rcGH standards ranging from 0-
400ng/ml were prepared and standard curve was made. Assay parameters like pH, temperature, reaction time, antibody
dilution and conjugate were optimized. The assay was simple to perform and was reliable. To determine the functional
activity of rcGH, receptor binding assay was optimized. rcGH was iodinated and its biological activity was detected by
using caprine liver receptor membrane and was dependent on receptor protein concentration, tracer counts, temperature,
time of incubation and assay pH. In cross-reactive study, human GH competed and displaced the iodinated rcGH by
binding effectively to the caprine receptor. Scatchard analysis of the rcGH binding suggested a single class of binding
site to the caprine liver membrane with an affinity of 337.1 ± 82.94 × 109 M-1 and capacity of 57.61 ± 4.23 fmol mg-1.

GENETICALLY SHAPING MORPHOLOGY OF THE FILAMENTOUS FUNGUS ASPERGILLUS
GLAUCUS FOR PRODUCTION OF ANTITUMOR POLYKETIDE ASPERGIOLIDE A

School of Biotechnology, East China University of Science and Technology, China

Abstract

Background: For filamentous fungi, the basic growth unit of hyphae usually makes it sensitive
to shear stress which is generated from mechanical force and dynamic fluid in bioreactor, and it
severely decreases microbial productions. The conventional strategies against shear-sensitive
conundrum in fungal fermentation usually focus on adapting agitation, impeller type and
bioreactor configuration, which brings high cost and tough work in industry. This study aims to genetically shape
shear resistant morphology of shear-sensitive filamentous fungus Aspergillus glaucus to make it adapt to
bioreactor so as to establish an efficient fermentation process.

Results: Hyphal morphology shaping by modifying polarized growth genes of A. glaucus was applied to reduce
its shear-sensitivity and enhance aspergiolide A production. Degenerate PCR and genome walking were used to
obtain polarized growth genes AgkipA and AgteaR, followed by construction of gene-deficient mutants by
homologous integration of double crossover. Deletion of both genes caused meandering hyphae, for which,
ΔAgkipA led to small but intense curves comparing with ΔAgteaR by morphology analysis. The germination of a
second germ tube from conidiospore of the mutants became random while colony growth and development almost
maintained the same. Morphology of ΔAgkipA and ΔAgteaR mutants turned to be compact pellet and loose clump
in liquid culture, respectively. The curved hyphae of both mutants showed no remarkably resistant to glass bead
grinding comparing with the wild type strain. However, they generated greatly different broth rheology which
further caused growth and metabolism variations in bioreactor fermentations. By forming pellets, the ΔAgkipA
mutant created a tank environment with low-viscosity, low shear stress and high dissolved oxygen tension, leading
to high production of aspergiolide A (121.7±2.3 mg/L), which was 82.2% higher than the wild type.

Conclusions: A new strategy for shaping fungal morphology by modifying polarized growth genes was applied in
submerged fermentation in bioreactor. This work provides useful information of shaping fungal morphology for
submerged fermentation by genetically modification, which could be valuable for morphology improvement of
industrial filamentous fungi.

Abstract

During assembly, HIV-1 must select its genomic RNA (gRNA) from a variety of cellular and viral
spliced RNAs. Despite a large number of studies, there is no consensus on how the Pr55Gag
precursor achieves this selection. These studies were limited by the expression and purification of
intact full-length Pr55Gag protein. Here, we purified soluble full-length Pr55Gag and we
investigated the specific determinants of the selective binding of Pr55Gag to HIV-1 gRNA using RNA binding and
footprinting assays. Our results revealed that Pr55Gag exhibits a higher binding affinity for gRNA than for spliced
vRNA species. Importantly, we demonstrate that the primary Pr55Gag binding site consists of the internal loop and the
lower part of stem-loop 1 (SL1), the upper part of which initiates gRNA dimerization. Further analyses on viral RNA
fragments of different length spanning the Psi and/or its flanking regions are in favor of a long-distance tertiary
interaction involving sequences upstream of SL1 and downstream of SL4, which promotes the optimal binding of
Pr55Gag to gRNA. Altogether our data shed light on the importance of a proper gRNA conformation that regulates its
specific binding to Pr55Gag, and could result in the competent selection and packaging of the genome. We propose a
new model to explain how Pr55Gag discriminates and specifically selects gRNA from cellular RNAs and viral spliced
vRNAs that also harbor functional SL1 in their first common exon. A double regulation ensures specific binding of
Pr55Gag to the gRNA despite the fact that SL1 is also present in spliced viral RNAs. The region upstream of SL1, which
is present in all viral RNAs, prevents binding to SL1, but this negative effect is counteracted by sequences downstream
of SL4, which are unique to the gRNA.

CONFERENCE ABSTRACT (SESSION)

Medical Biotechnology (Track)

SURVIVIN EXPRESSION IN RENAL CELL CARCINOMA AND ITS CORRELATION WITH
CLINICOPATHOLOGICAL PARAMETERS

Khaled El Gehani

Department of Pathology, University of Benghazi, Qar Yunis, Benghazi, Libya

Abstract

Objective: The aim of the current study is to cast further light on the issues related to prognostication
of renal cell carcinoma (RCC), assessing the expression of survivin in a subset of primary RCC and
determine its relation to different clinicopathological features and disease free survival. Methods: The
present series consisted of tissue samples obtained from 37 Libyan patients with stage I, II, III, or IV
RCC. Survivin expression in these tumors was assessed by immunohistochemistry using an automated staining system.
Different grading systems were tested for expression of survivin.

Conclusion: Survivin expression in RCC may identify patients at risk of a more aggressive disease and a worse
prognosis, further investigations, on a larger and more heterogeneous population, should be carried out to validate and
extend our results.

Abstract

AGEs are a heterogeneous group of compounds derived from non-enzymatic glycation of proteins,
lipids and nuclic acids through complex reaction known as the Maillard reaction. AGEs interact with
the receptor for AGEs (RAGE) on the membrane and induce harmfull effects through activation of
nuclear factor kappa-B, and increased oxidative stress and inflammatory mediators. AGEs combine with membrane
receptors (RAGEs) also combine with circulating soluble receptors (sRAGE) and act as a decay agent. AGEs are
thought to be involved in many complications of angiopathy, cardiovascular diseases (CVD) and nephropathy in
diabetes as well as those of end‐stage renal failure. The aim of this study was to evaluate the changes of (AGEs),
(sRAGE) and Oxidized Low-density Lipoproteins ( OxLDL) levels, in hemodialysis patients with different underlying
causes and to evaluate them in relation to occurrence of CVD or with diabetic nephropathy as an underlying cause in
relation to other glycemic, lipid profile and renal functions' biomarkers.Our study included 279 patients with end stage
renal disease (ERD) who received maintenance hemodialysis (HD) (duration of HD, 6.7 ± 3.4 years.) and 112 sex and
age matched healthy control subjects.The underlying casual disorders for them were as follows :diabetic nephropathy
(56.9%) and non-diabetic nephropathy diseases (43.1%) including chronic hypertensive nephrosclerosis,
glomerulonephritis, polycystic kidney disease, and rest of unknown etiology. We also categorized HD patients into 2
subgroups according to their positive medical history of CVD ( 52.09%) and another subgroups with no medical history
of CVD (47,01%).We demonstrated significantly higher levels of oxLDL, AGES and sRAGE in HD than healthy
control group ( P< 0.001). We found significant increase of OxLDL, AGEs and sRAGE in HD diabetic nephropathy
subgroup when compared with non-diabetic subgroup as an underlying cause. Interestingly sRAGEs significantly
decreased in HD patients who had positive history for CVD in comparison with those HD patients with no history of
CVD (1970 ± 870, 2660 ± 908; P < 0.001). There was positive correlation between AGEs and sRAGE in whole HD
patients (r =0.441;P = < 0.001, r = 0.395; P= < 0.05) while only in HD group with CVD There was negative correlation
( r = - 0.294 ; p = <0.01). We found that AGEs was an independent determine for diabetic nephropathy as an underlying
cause for ESR (OR, 1.25; 95% CI, 1.04 to 1.49; P < 0.016). Another factor was the oxLDL level (OR, 1.10; 95% CI,
1.02 to 1.19, P < 0.018) and sRAGE (OR, 1.04; 95% CI, 1.01 to 1.06, P < 0.007). and we identified sRAGE as an
independent factor associated with the prevalence of CVD(OR, -0. 49; 95% CI, -0.086 to 0.034; P < 0.001). The other
factors were AGEs (OR, 2.81; 95% CI, 1.79 to 4.41; P < 0.001), ox LDL (OR, 1.04; 95% CI, 1.02 to 1.06; P <
0.001),and hs-CRP level (OR, 1.02; 95% CI, 1.00 to 1.05, P <0.046).Conclusions plasma OxLDL,AGEs and sRAGE
levels are strongly associated with the prevalence of cardiovascular disease and diabetic nephropathy in hemodialysis
patients and could be considered future markers for diabetic nephropathy and cardiovascular diseases in hemodialysis
patients with end stage renal disease.

BLOCKING THE FUNCTION OF INFLAMMATORY CYTOKINES AND MEDIATORS BY
USING IL-10 AND TGF-Β: A POTENTIAL BIOLOGICAL IMMUNOTHERAPY FOR
INTERVERTEBRAL DISC DEGENERATION IN A BEAGLE MODEL

Abstract

Introduction: The debilitating effects of lower back pain are a major health issue worldwide. A
variety of factors contribute to this, and oftentimes intervertebral disk degeneration (IDD) is an
underlying cause of this disorder. Inflammation contributes to IDD, and inflammatory cytokines play
key roles in the pathology of IDD. This study characterized the potential to suppress inflammatory
cytokine production in degenerative intervertebral disc (NP) cells by treatment with IL-10 and TGF-β in a canine model
of IDD.

Methods: IDD was induced surgically in six male beagles, and degenerative NP cells were isolated and cultured for in
vitro studies on cytokine production. Cultured degenerative NP cells were divided into four experimental treatment
groups: untreated control, IL-10-treated, TGF-β-treated, and IL-10- plus TGF-β-treated cells. Cultured normal NP cells
served as a control group. TNF-α expression was evaluated by FACS analysis and ELISA; moreover, ELISA and realtime
PCR were also performed to evaluate the effect of IL-10 and TGF-β on NP cell cytokine expression in vitro.

Results: The major findings of these analysis are that after treatment with IL-10 and TGF-β, the expression of
extracellular and intracellular TNF-α and IL-1β was suppressed, while the expression of inflammatory cytokines in
untreated normal NP cells was significantly lower than that in untreated degenerative NP cells. Our results demonstrated
that IL-10 and TGF-β treatment suppressed the expression of IL-1β and TNF-α and inhibited the development of
inflammatory responses.

Discussion: We observed that either TGF-β or IL-10 alone suppressed the expression of inflammatory cytokines.
Furthermore, their combined use produced a higher level of inhibition of TNF-α and IL-1β than either TGF-β or IL-10
alone. IL-10 and TGF-β should be evaluated as therapeutic approaches for the treatment of lower back pain mediated by
IDD.

CONFERENCE ABSTRACT (SESSION)

Medical Biotechnology

HIGH RAB27A EXPRESSION INDICATES FAVORABLE PROGNOSIS IN CRC

Pingsheng Chen and Shi Chuan Bing

Department of Pathology and Pathophysiology, School of medicine, Southeast University, Nanjing,
China

Abstract

Background: Rab27A is a peculiar member in Rab family for its specific role in human hereditary
disease. Recently, Rab27A has been suggested to play essential roles in the development of several
kinds of human cancers. However, the association between Rab27A expression and
clinicopathological characteristics of colorectal cancer (CRC) has not been elucidated yet.

Methods: One-step quantitative real-time polymerase chain reaction (qPCR) test with 18 fresh-frozen CRC samples and
immunohistochemistry (IHC) analysis in 112 CRC cases were executed to evaluate the relationship between Rab27A
expression and the clinicopathological features of CRC. Cox regression and Kaplan-Meier survival analyses were
performed to identify the possible factors which correlated with the prognosis of CRC patients.

Conclusion: The data indicated the differentiate expression of Rab27A in CRC tissues and matched non-cancerous
tissues. Rab27A may be used as a valuable prognostic biomarker for CRC and that high Rab27A expression suggests a
favorable prognosis for CRC patients.

BRING SANGER SEQUENCING TO THE COMMUNITY CLINICAL LABORATORIES
WORLDWIDE

Abstract

One obstacle to implement automated Sanger sequencing as routine tests in developed countries and
as needed accurate diagnostic tools for emerging infectious diseases, such as Ebola, is the labile PCR
reagents, namely the DNA polymerases, the dNTPs and the reverse transcriptases, which must be
kept at -20°C between uses. We have optimized a PCR chemical mix in which these labile
components are stabilized at room temperature for several weeks to 10 months. It depends on a highly processive,
moderately heat-resistant DNA polymerase with a PCR thermocycling not to exceed 85°C [1], and has been successfully
used for preparing DNA sequencing templates in the routine detection and genotyping of human papillomaviruses [2], in
the diagnosis of Neisseria gonorrhoeae and Chlamydia trachomatis [3] and in the diagnosis of Lyme borrelioses [4-6].
By adding a reverse transcriptase in this low temperature PCR mix, the newly created RT-PCR mix may be adapted for
screening Ebola patient samples at the sites of outbreak in far-flung places and the PCR amplicons can be transported to
a regional laboratory for DNA sequencing validation.

Abstract

Glycosylation is a form of co-translational and post-translational modification of proteins.
Glycans have a variety of structural and functional roles in membrane and secreted proteins.
Changes in glycosylation are accompanied with changes in physiological state, which are also
associated with various diseases such as cancer, AIDS, rheumatoid arthritis, multiple sclerosis
and many others. Glycoprofiling thus could serve as a diagnostics tool for even early stage of diseases. A highly
challenging is the fact that analytical detection platform for glycan analysis has to be able to detect very low
levels of analytes. The aim of this work was to develop lectin-based protein microarray method for glycoprofiling
of proteins and to apply this method for analyses of glycoproteins isolated from the samples by
immunoprecipitation. We measured proteins and receptors of insulin-like growth factor (IGF) system isolated
from serum, cytosol or membrane protein fraction of two groups of the samples, (1) serum of healthy persons and
patients with colorectal cancer and healthy and tumor part of the tissue of the same patient, and (2) placental
membranes from healthy mothers, mothers with pre-eclampsia, and mothers with diabetes. The microarray slides
with spotted samples were incubated with set of biotinylated lectins with various glyco-specificity, labeled with
streptavidin-fluorescent dye conjugate and scanned. The appropriateness of developed lectin-based protein
microarray method for biosensing of changes in protein glycosylation of IGF system was evaluated, and the
differences in protein glycosylation within each sample group (for colorectal cancer healthy vs. tumor tissue, for
placental samples healthy vs. pre-eclampsia vs. diabetes) were quantified.

ACKNOWLEDGEMENT
This work is supported by the grants: VEGA 2/0162/14 and APVV 0282-11.

Medical Biotechnology (Track)

APOPTOTIC EVENTS INDUCED BY PROTOTYPE FOAMY VIRUS INFECTION

Abstract

Foamy virus infection induces cytopathology in several cell types from different species. The
exact mechanism of cell killing by foamy viruses is still unknown. In this study, we have
investigated the mechanism of cell death induced by prototype foamy virus (PFV) infection in
baby hamster kidney (BHK 21) cells lines. PFV induces apoptosis by exhibiting morphological
alterations such as chromatin condensation, blebbing, and nuclear fragmentation. In addition, PFV infection
causes the fragmentation of chromosomal DNA, upregulation of Bax, and activation of caspases-3. Upregulation
of Bax initiates the translocation of cytochrome-c from mitochondria to the cytoplasm, suggesting that PFVinduced
apoptosis is triggered predominantly via the mitochondrial mediated pathway. Blocking apoptosis using
caspases inhibitors increased PFV-infected BHK 21 cell viability. Although blocking apoptosis resulted in
reduced progeny release, maximal accumulation of PFV was found in apoptosis-blocked cells. This report
provides the first systematic experimental account of the mechanism of apoptosis induced by PFV infection,
which will provide valuable insights for understanding why pathology is not associated in animals infected with
cytopathic foamy viruses.

Clinical Research Center, National Hospital Organization, Sagamihara National Hospital, Japan

Abstract

Rationale: To investigate the role of helper T (Th) cells in steroid resistant (SR) asthma, steroid
sensitive (SS) and resistant (SR) Th clones were selected in vitro, and then adoptively transferred
into unprimed mice. Effect of CTLA4-Ig was analyzed both in vitro and in vivo.

Methods: For in vitro evaluation, ovalbumin (OVA) reactive Th clones were cultured with antigen
presenting cells and OVA in the presence of various concentrations of dexamethasone (DEX). Proliferative responses of
Th clones were measured by 3H-thymidine incorporation. For in vivo assessments, unprimed BALB/c mice were
transferred with Th clones, challenged with OVA, and administered with DEX subcutaneously. Bronchoalveolar lavage
fluid (BALF) was obtained 48 hr after challenge, and the number of infiltrating cells was differentially counted. CTLA4-
Ig was administered through nasal inhalation or venous injection.

Results: SS and SR clones were selected based on the effect of DEX on the proliferative responses of antigen-stimulated
Th clones. Airway infiltration of eosinophils and lymphocytes of mice transferred with SS clones were effectively
inhibited by the administration of DEX. In contrast, those of mice transferred with SR clones were not significantly
inhibited by DEX. Administration of CTLA4-Ig significantly suppressed the proliferation of DEX-treated SR clones in
vitro, and the eosinophil infiltration of SR asthma model transferred with SR clones in vivo.

Conclusions: Steroid sensitivity of Th clones assessed in vitro was consistent with that of adoptively transferred asthma
model assessed in vivo. Costimulatory signal mediated through CD28 is crucial for the induction of steroid resistance
both in vitro and in vivo.

Methods and Materials: A total of 422 ears from 211 patients with HNC were included in the study. The analytical
endpoint was defined as grade 2+ tinnitus that occurred within 1 year of IMRT. The logistic and LΚΒ NTCP models
were used to assess the incidence of grade 2+ tinnitus. The best-fit values for TD50 were determined using maximumlikelihood
estimates. A guideline of TD25 is recommended for the tolerance dose to produce a 25% complication rate
within a specific period of time.

Abstract

Methods: In total, 120 patients with breast cancer were analyzed. All participants were treated with
IMRT or hybrid IMRT. The final endpoint was defined as those who with symptomatic pneumonitis
combined with computerized tomography (CT) measured density changes grade ≥1. The risk factors for a multivariate
logistic regression normal tissue complication probability (NTCP) model of SRP were determined using the least
absolute shrinkage and selection operator (LASSO) technique.

Results: Three risk factors were selected using LASSO: the percentage of the ipsilateral lung volume that received more
than 20-Gy (IV20), age, and body mass index (BMI). Our analyses indicate that the risk of SPR following IMRT/hybrid
IMRT in elderly or low-BMI breast cancer patients increases when the percentage of the ipsilateral lung volume
receiving more than 20-Gy is controlled below 30%.

Conclusions: We suggest to define a dose-volume percentage constraint of IV20Gy < 30% for the irradiated ipsilateral
lung in radiation therapy treatment planning to maintain the incidence of SPR below 10%, and pay attention to the
sequelae especially in elderly or low-BMI breast cancer patients.

A NEW ASSAY FOR MEASURING TRACE ELEMENTS IN SERUM FOR THE ESTIMATION OF
BREAST CANCER RISK

Abstract

An assay to determine the concentrations of trace elements in serum can reflect the metabolic status
of the whole body, as trace elements are linked to all of the body's metabolic systems as catalysts.
Such an assay could also be measured simply, without imposing physical burdens on subjects. Our
group investigated the balance of trace elements in serum to elucidate the correlations between trace
elements and cancers. We then used our data on the correlation to develop a new method of
measuring trace elements in serum. Venous blood samples were collected from Japanese breast cancer patients before
medical treatment (n=30), and from age-matched controls undergoing medical examinations (n=30). After serum
separation, 16 elements (Na, Mg, Al, P, K, Ca, Ti, Mn, Fe, Cu, Zn, Se, Rb, Ag, Sn, S) were measured by ICP-MS. The
"metallo-balance" determined by a multivariate analysis of the trace element profiles of the two groups was effective
marker for breast cancer risk. The ROC curve calculated to evaluate performance had an AUC of 0.999. Multivariate
discriminating classifiers based on trace element profiles measured by ICP-MS were successful in discriminating breast
cancer. Our results suggest that serum trace element profiling has great potential for the diagnosis of breast cancer.

NEW PERSPECTIVES IN NANOMEDICINE AND NANOPHARMACOLOGY

Abstract

The strategic combination of various biophysical and biochemical methods such as NMR, X-ray
crystallography, Atomic Force Microscopy, Accelerated Mass Spectrometry and Surface Plasmon
Resonance techniques with cell biological approaches and patient studies leads to new perspectives in
the fields of nanomedicine and nanopharmacology concerning innovative routes in applied health
care. These routes show solutions for different so-far unsolved medical and pharmacological problems in oncology, antiinfection,
tissue-engineering and pathobiochemistry of the endocrine system [1-7]. It was essential for this purpose to
decipher the three alphabets of life completely (1: nucleic acid code, 2: amino acid code, 3: sugar code [8]) and figure
out how they work together on a sub-molecular level. It turned out in our molecular dynamics simulations that ab initio
calculations play an important role in order to fit all the results from the different approaches together in a convincing
way.

STRUCTURE AND STABILITY OF A MEMBRANE PROTEIN:AMPHIPOL VACCINE
FORMULATION

Abstract

We have developed a vaccine formulation based on a purified membrane protein solubilized in an
amphipathic polymer (amphipol, APol). We have chosen Chlamydia trachomatis as a target for
vaccine development; C. trachomatis is a major bacterial pathogen throughout the world. Efforts
have focused on the production of a subunit vaccine directed against C. trachomatis major outer
membrane protein (MOMP). MOMP is purified in its native (n) trimeric form using the zwitterionic detergent Z3-14,
but its stability in detergent solutions is limited. Amphipols (APols) are synthetic polymers that can stabilize membrane
proteins (MPs) in detergent-free aqueous solutions. APols maintain nMOMP secondary structure and that
nMOMP/APol vaccines elicit better protection than formulations using nMOMP solubilized in Z3-14. We used
spectroscopic techniques to characterize the secondary structure (circular dichroism), tertiary and quaternary structures
(immunochemistry and gel electrophoresis) and aggregation state (light scattering) of nMOMP in APols as a function of
temperature and time. Our analyses show that APols protect nMOMP much better than Z3-14 against denaturation due
to continuous heating, repeated freeze/thaw cycles, or storage at room temperature. We envision a future where classical
vaccines will be replaced by purified membrane protein/Apol vaccines that are more effective, safer, and do not require
refrigeration.

Abstract

The effect of ethanol extract of cyperus rotundus rhizome on carbimazole - induced hyperlipidemia
and on hyperglycemia (using glucose tolerance test) in male wistar rats was investigated. Acute
toxicity analysis with the cyperus rotundus extract produced no lethality even at high doses.
Hyperlipidemia was induced using 400mg/kg cholesterol and 2mg/kg carbimazole. The lipemic
control groups were administered cholesterol and carbimazole but not the normal control group.
Cholesterol and carbimazole administration caused a significant (p<0.05) increase in the Total Cholesterol, Triglyceride
(TG), Low Density Lipoprotein (LDL), Non-High Density Lipoprotein (non-HDL) Cholesterol and LDL/HDL ratio and
a significant (p<0.05) decrease in the levels of HDL in the lipemic control when compared to the normal control.
Treatment with cyperus rotundus extract at 250mg/kg, 500mg/kg and the standard hyperlipidemic drug (simvastatin)
5mg/kg significantly (p<0.05) reduced Total Cholesterol, TG, LDL, LDL/HDL ratio, Total non-HDL Cholesterol and
significantly (p<0.05) increased the level of HDL when compared to the non-treatment groups (the normal control and
the lipemic control). Treatment with cyperus rotundus extract and the hyperglycemic drug (Glucophage) at 7.14mg/kg
caused a significant (p<0.05) decrease in the blood glucose level when compared to the non-treatment groups. This study
concluded that cyperus rotundus rhizome contains principles that compare effectively and as well as standard clinically
used antihyperlipidemic and antihyperglycemic agents.

Abstract

The biospecificity of antibody – antigen (Ab–Ag) agglutination is just one example of immense
variety of intricate protein-protein interactions. These interactions include different interplay of
forces, which explain mechanical, electrical and thermal energies that form the internal system
energy. The method of measuring of internal energy of serological agglutination suspension has been
approved by FIPS RU [1]. The physical aspects of protein-protein interaction on the model of serological
immunosuspention tests on the base of latex agglutination (LA) and red blood cells (ram’s erythrocytes)
hemagglutination (HA) tests were first established in SI units [2, 3]. Despite the fact that the first time this phenomenon
was observed, it was described as an affinity; in fact, it’s the real mechanical part of internal energy in the serological
system.

On the basis of differences in potential energy in positive and negative results (“umbrella” and “button”) in agglutination
tests and measuring the mass of carries (latex particles, red blood cells), we have calculated the force and energy of
biospecific interaction of active centers in Ab–Ag reaction in Newtons and Joules. We have used in our investigation
organic and native ingredients with non-standardized properties.

The model of Ab-Ag interaction is the most convenient for measuring in physical units the biospecific and non-specific
protein molecules interactions. The correlation when using different antigen’s diagnostics as for plague, brucellosis and
tularemia infections which are nearly ten times differ in sensitivity have shown the same deviations in their force and
energy. Thus, antibodies to the causative agent of plague comparing with those of brucellosis and tularemia showed data
1,03×10-17, 5,1×10-17 and 1,1×10-18 of Newton per single molecule of antibody, respectively. The energy data of
plague antibody exceeded up to 10 times those above mentioned.

The usage of standardized latex particles and synbodies for artificial calibrated test system on account of sensitivity and
stability would permit us to create physical biospecificity unit for SI system. This creation may help us to develop and to
take under control the up-to-date idea concerning the quantum coherence electromagnetic waves signal, their energy and
molecules distance recognition that will be primarily used in the investigation of protein molecules interaction and their
signaling, transduction systems of signal and practical utilization.

CONFERENCE ABSTRACT (SESSION)

Pharmaceutical Biotechnology

ENDOPHYTIC FUNGUS AS A SOURCE OF BIOACTIVE SECONDARY METABOLITES:
RESVERATROL AND VINIFERIN

Chetana Roat and Shailesh R. Dave

Microbiology and Biotechnology, School of Sciences,Gujarat University, Ahmedabad, India

Abstract

Resveratrol and Viniferin possess a broad spectrum of pharmacological and therapeutic effect like
anti-cancer, antioxidant, anti-atherosclerosis. Endophytes, by residing within the specific chemical
environment of host plant tissue, form unique group of microorganisms. Microbially unexplored
medicinal plants can have diverse and potential microbial association. The aim of this work is to
identify and isolate the endophytic microorganisms that possess resveratrol and viniferin producing capability and
optimizes the conditions for their production. Endophytes from Vitis vinifera L.cv. Merlot, Vitis quinquangularis Rend.
and Cayratia trifolia L. were studied and the resveratrol and viniferin producing isolates were screened by thin Layer
Chromatographic (TLC) with solvents system CHCl3/MeOH/HCOOH, 85/15/3 (v/v) and identification of the compound
was done by HPLC. The total of 20 isolates were obtained, 5 were endophytic bacteria and 15 were endophytic fungi
and it was found that the one strain Aspergillus sp. AB4 isolated from leaf of Vitis vinifera using Czapek yeast extract
agar medium had stable resveratrol and viniferin producing capability in all subcultures. Cell growth of Aspergillus sp.
increased during cultivation and reached a stable and high level of biomass after 7 days. The best fermentation
conditions for resveratrol and viniferin production in broth cultures of Aspergillus sp. AB4 were an inoculum size of
10%, a rotation speed of 100 rpm , and a temperature of 28°C.

CONFERENCE ABSTRACT (SESSION)

Pharmaceutical Biotechnology

LONG ACTING ANALOGS OF GLYCOPROTEIN HORMONES DESIGNED BY SITE-DIRECTED MUTAGENESIS AND GENE FUSION

Fuad Fares

Department of HumanBiology, Faculty of Natural Sciences, University of Haifa, and Department of Molecular Genetics, Carmel Medical Center, Haifa, Israel

Abstract

One major issue regarding the clinical use of many peptides is their short half-life due to the rapid clearance from the circulation. To overcome this problem, we used overlapping PCR technique to add the signal sequence of O-linked oligosaccharides to the coding sequence of glycoprotein hormones. The used cassette gene contains the sequence of the carboxyl-terminal peptide (CTP) of human chorionic gonadotropin β (hCGβ) subunit. It was postulated that the O-linked oligosaccharides add flexibility, hydrophilicity, stability and prevent plasma clearance and thus increasing the half-life of the protein in circulation. Using this strategy we succeeded to ligate the CTP to the coding sequence of follitropin (FSH), thyrotropin (TSH), erythropoietin (EPO) growth hormone (GH) and thus to increase the longevity and bioactivity of these proteins in-vivo. Interestingly, the new analog of FSH was found not immunogenic in humans and it was approved by the European Commission (EC) for treatment of fertility. In addition, our results indicated that long acting GH is not toxic in monkeys and the results from clinical trials phases I and II seem to be promising. Designing long acting peptides will diminish the cost of these drugs and perhaps reduce the number of injections in the clinical protocols.

Deletion of the N-linked oligosaccharides from hTSH significantly reduced its activity in vitro and in vivo. Moreover, deglycosylated TSH compete with hTSH and human Thyroid Stimulating immunoglobulin (TSI) in a dose dependent manner. These variants may offer a novel therapeutic strategy in the treatment of hyperthyroidism and Grave’s disease.

Abstract

To bridge the GAP between basic and applied sciences is needed to push forward disease research
and therapeutics. What types of drug leads are truly “druggable”, sit in “patented bioproducts space”
and can be pushed towards clinical trials? We present two successful translational cases of
bioproducts from laboratory bench to the bedside.

Case 1: Fibrin sealant derived from snake venom (FSSV). This product is composed of a serine protease (thrombin-like
enzyme) derived from South American rattlesnake (Crotalus durissus terrificus) venom and cryoprecipitate of buffaloes
(Bubalus bubalis) rich in fibrinogen. The activity of snake venom serine protease is 1,500-fold more potent than that of
human thrombin employed in commercial sealants. The cryoprecipitate replaces human hemoderivatives, which makes
the production more affordable and eliminates the possibility of infectious disease transmission. FSSV is a bioproduct of
animal origin produced at the Center for the Study of Venoms and Venomous Animals (CEVAP) of UNESP, Brazil, to
treat chronic venous ulcers of 240 patients of a phase II/III multicenter clinical trial. The phase I/II clinical trial with ten
pacients was carried out and there was complete healing in 38.8% of the ulcers. There was decrease of their area with
wound bed preparation in 33.3%, therefore a significant improvement in 72.1% of the cases was observed in only three
months of treatment. Based on such results, the product was considered a safe candidate and clinically promising. In
addition, FSSV is being successfully tested in vitro and in vivo for its use as a three-dimensional scaffold for stem cells,
since it is biodegradable and maintains the cells viable at the application site.

Case 2: Africanized honeybee antivenom (AHBA). In Brazil, Africanized honeybees provoke more than 10 thousand
envenomations and approximately 40 deaths per year. Since it is an issue limited to the American continent that had no
specific treatment, a new antivenom to treat multiple stings was developed. One of the major challenges was the
standardization of the product, because of the antigenic characteristics of the venom. The antivenom was developed at
CEVAP and produced by the Vital Brazil Institute (IVB) based on horse hyperimmunization with the main toxic
fractions of the venom. Each milliliter of AHBA neutralizes 1.25 mg of venom and 10 mL of antivenom can treat about
100 stings. AHBA is being tested in a phase I/II multicenter clinical trial with 20 participants. Its delivery route is
intravenous and the administration follows a strict clinical protocol that aims to rapidly neutralize and eliminate the
venom toxic effects.

Both products were patented and by the end of the clinical trials, they will be available in the Brazilian public health
system (SUS). They will aid the treatment of chronic venous ulcer patients and multiple Africanized Honeybee sting
victims, two major neglected issues. The involved team of researchers acquired the required expertise to develop new
drugs through translational research by applying modern biotechnological tools and aiming at solving health problems
with regional bioproducts.

THE EVALUATION OF CERCARIAL TRANSFORMATION FLUID USED SINGLY AND IN COMBINATION WITH CRUDE CERCARIAL ANTIGEN IN EXPERIMENTAL SCHISTOSOMIASISMANSONI.

Abstract

Schistosomiasis is a truly neglected tropical disease and the second major parasitic disease in the world after malaria. It affects 201.5 million cases in Africa alone. The goal of this research is the development of an appropriate vaccine against experimental schistosomiasismansoni, we assessed the effect of cercarial transformation fluid (CTF) singly and in combination with crude cercarial antigen (CCA) using alum as an adjuvant in experimentally infected mice. The combined antigen gave the best results as evidenced by significant reduction in the worm load (62.07%), tissue egg count, (78.16%, 86.46%) in liver and intestine respectively and hepatic granuloma size (31.76%). Scanning electron microscopic study revealed changes in the tegument in the form of roughness and appearance of vesicles and furrows between the tegumental tubercles. Also, resorption and mutilation of the ventral sucker and dimples replacing its spines. The female tegument was irregular and its posterior end showed loss of spines and sensory bulbs. Moreover there was a significant decrease in liver enzymes (ALT and AST) compared to infected control mice. A significant elevation in CD4+ T-lymphocytes denoting amelioration of the immune status in animals received combined vaccine.

It can be concluded that this combined antigen give us a hope to the development of a feasible simple vaccine against Schistosomiasismansoni.

Xenobiotics Research Unit, The POntifical catholic University of Paraná, Curitiba, Brazil

Abstract

Microbial biotransformation (a.k.a. biocatalysis, bioconversion) processes are amongst the smarter
and greener ways to obtain new therapeutic molecules. Such processes occur when a microorganism
converts a substrate into new molecule(s) by adding chemical functional groups, mainly via P450
cytochromes. Consolidated data reveals that immobilized cells tend to carry out biotransformations in
a more promptly manner and several processes have been developed for that. In this study, we propose the use of filter
paper as a support matrix for microbial attachment and further growth as biofilm.

A dedicated biofilm bioreactor was conceived and built. Among a diversity of microorganisms possible to be employed,
Candida albicans was elected due to its wide morphological multiplicity. Eight-millimeter paper disks were soaked with
ca. 106 cells of C. albicans and transferred to the biofilm bioreactor. Sterile culture broth was admitted in a slow flux
and consistent biofilms were obtained after 72 h. Scanning electron microscopy revealed the occurrence of pseudo and
true hyphae in abundance, even within the paper matrix.

Authors propose that paper-based matrixes may be considered as suitable supports for the growth of biofilms, which
may serve as living biocatalysts.

POSTER PRESENTER

CONFERENCE ABSTRACT (POSTER)

Pharmaceutical Biotechnology (Track)

ENGINEERING OF CYTOKINE GENE FOR CONDITIONAL ACTIVATION

Iram Gull, Muhammad Shahbaz Aslam and Muhammad Amin Athar

Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan

Abstract

Use of recombinant cytokines for therapeutic purposes has side effects due to the pleiotropic effect. It
diminishes the efficacy of recombinant cytokines. Use of therapeutics in the form of pro-drug can
eliminate the chances of non-specific binding and subsequent adverse effects. This approach
ultimately improves the drug efficacy and increases the trust of consumer on therapeutic cytokines for
safe use. In the present study, the gene of interferon alpha 2b has been engineered by fusion with gene of latency
associated protein (LAP) of human TGFβ using HCV NS3 protease cleavage site as linker for conditional activation of
resultant chimeric protein only in HCV infected cells by positive use of hepatitis C virus NS3 protease activity.

IN VIVO ANTI-HYPERGLYCEMIC AND ANTIOXIDANT ACTIVITIES OF THE LEAF
EXTRACTS OF HOLARRHENA ANTIDYSENTERICA ON ALLOXAN-INDUCED DIABETIC
RATS

Abstract

Medicinal plants constitute an important natural wealth of a country. They play a significant role in
providing primary health care services to rural people. They serve as therapeutic agents as well as
important raw materials for the manufacture of traditional and modern medicine. The present study
was undertaken to evaluate the anti-hyperglycemic activity of the crude extracts of leaves of
Holarrhena antidysenterica. The pet ether, chloroform and ethanolic extracts have been subjected to estimate the antihyperglycemic
activity in alloxan-induced diabetic rats. Blood glucose levels were measured using the commercially
available glucometer. Glibenclamide was used as a reference drug at a dose of 0.6 mg/kg. The antioxidant activity of the
extracts has been studied in the liver tissue of diabetic rats by measuring catalase and lipid peroxidation levels. The
results showed that, ethanolic extract possessed a significant anti-hyperglycemic and antioxidant activity, which is
equipotent with the reference drug.

LAVANDULA OFFICINALIS ESSENCIAL OIL AGAINST HAEMONCHUS CONTORTUS

Abstract

The sheep farming is traditionally practiced worldwide as a promising economic activity. However,
the general unpreparedness of the breeders in the management of animals has contributing to the
development of gastrointestinal infections mainly caused by Haemonchus contortus species. H.
contortus sheep infections have been related to serious economic losses on sheep farming and the
irregular or erroneous administration of traditional anthelmintic drugs has been associated to the resistance development.
Ethnopharmacological information has appointed the Lavandula officinalis essential oil as a promising alternative for
development of new anthelmintic products. Thus, the main aim of this work was to investigate the popular use of
essential oil of L. officinalis with emphasis on its potential anthelmintic effects against H. contortus. According our best
results, L. officinalis essential oil showed 92.3% and 40.6% of efficacy in egg hatch and larval motility tests,
respectively. In the adult worm motility test, 85% of worms were completely immobilized within the first 6 hours of
nematode exposition to different dilutions of oil. Phytochemical analysis revealed the presence of linalool (36.01%),
sabinene (31.1%) and 1,8-cineol (5.61%) as the most important compounds, which may be responsible for anthelmintic
essential oil activity.

Pharmaceutical Biotechnology (Track)

Abstract

Modern civilization is facing hundreds of disorders associated with free radicals. The natural
antioxidants from non-edible plants are gaining importance to fight against these disorders. The
purpose of this study is to evaluate the antioxidant and antitubercular activities of Leucas
marrubioides Desf. (Lamiaceae) root extracts of petroleum ether, chloroform and methanol. The
antioxidant activity of the extracts has been evaluated using DPPH radical scavenging, reducing power and nitric
oxide scavenging methods. The results of the study indicated that, methanol extract possess promising DPPH
radical scavenging, reducing power and nitric oxide scavenging activity. The antitubercular activity of all the
extracts of L. marrubioides have been evaluated against Mycobacterium tuberculosis H73Rv strain using
Microplate Alamar Blue Assay (MABA). The activity was documented within MIC range of 0.2 to 100μg/ml. The
results of MABA showed that petroleum ether extract exhibited excellent antitubercular activity. The present
investigation suggests that L.marrubioides possess remarkable antioxidant and antitubercular activity.

CONFERENCE ABSTRACT (POSTER)

Pharmaceutical Biotechnology (Track)

ROLE OF WOMEN STUDIES IN INHIBITORY EFFECTS OF GLORIOSA SUPERBA EXTRACT
ON RHEUMATOID ARTHERITIS

Veena Megaravalli Krishnappa Hegde and H. N. Girish

Department of Studies and Research in English, Sahyadri Arts and Commerce College, Shimoga,
577201, Karnataka, India

Abstract

Women in many developing countries are often the protectors of biodiversity and have a developed
understanding of the medicinal and nutritional uses of a plethora of rare wild and cultivated plants.
Biodiversity is fundamental for the continued growth, sustainability and vitality of individuals and
communities across the globe. Women, especially in developing nations, are most vulnerable to
change in biodiversity and at the same time most capable of protecting and retaining biodiversity. Here is a case study on
inhibitory effects of gloriosa superba extract on Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory disease affecting about 1 % of the population in developed
countries. Gloriosa superba Linn. (family-Liliaceae) is one of the herbaceous climbers distributed throughout Western
Ghats and well documented traditionally in Ayurveda system of medicine for various ailments like inflammation, gout,
gonorrhea, leprosy, rheumatoid arthritis, jaundice, etc. The plant is highly valued in modern medicine owing to the
presence of alkaloids. Despite its wide spread use in traditional medicine for treatment of gout and rheumatism, the
present investigation was to evaluate the effect of chloroform extract of tubers of Gloriosa superba in Freund's complete
Adjuvant (FCA) induced arthritis using albino rats. The chloroform extract of tubers of Gloriosa superba has shown a
dose dependent and significantly decreased paw edema and ankle diameter in treated groups as compared with arthritic
group. Synovial membrane damage and neutrophil infiltration in histopathological examination was restored
significantly by the extract as compared to arthritic group. Eco-feminism is a movement that applies feminist principles
and ideas to ecological issues such as women's rights, peace, labour, ecological and environmental justice. Moreover, the
slogan "glocal", that is from global to local, which emphasizes localized cultures, economies, community based
sustainable practices that need to be rejuvenated for a harmonious relation between nature, man and woman.

SESSION SPEAKERS

CONFERENCE ABSTRACT (SESSION)

Plant and Environment

A NEW PLANT OIL PRODUCTION PLATFORM: SEED-LIKE OIL YIELD FROM BIOMASS

James Petrie, Thomas Vanhercke, Allan Green and Surinder Singh

CSIRO Food, Nutrition and Bioproducts Flagship, ACT, Australia

Abstract

Supply of vegetable oils as a major commodity faces continuous pressure. Global demand is expected
to double in the next two decades due to increasing world population and rising petroleum prices.
Increasing limitations on arable land and agricultural inputs mean it will be difficult to meet this
additional demand with current oilseed-based production systems. The concept of producing oil in
the leaves and stems of high biomass species has attracted attention as a way to intensify oil
production. The engineering of such a new oil production platform would not only yield greater amount of oil for a
given land area but also provide a way to more easily segregate bioeconomy traits such as unusual fatty acids away from
food production. We previously reported the accumulation of up to 17% triacylglycerol (TAG, dry weight) in leaf tissue
of Nicotiana species. This was achieved by combinatorial metabolic engineering in which we increased fatty acid
biosynthesis (‘Push’) by limited overexpression of the WRI1 transcription factor, increased TAG assembly (‘Pull’) by
expressing DGAT1, and encouraged oil body formation (‘Packaging’) by expressing oleosin in plant leaves1,2.

In this presentation, we will describe some of our second generation construct designs which have more than doubled the
previously reported TAG content. Oil content in leaves now matches elite oilseed crop seed levels of +40%. We will
describe the implications that this technology has for global plant oil production from a yield and intensification
perspective, as well as the challenges that remain for integration into the existing industry. We will also present data
demonstrating that the newly produced fatty acids can be modified for industrial or nutritional applications as well as
preliminary data of a transcriptome comparison between wild type and high oil leaf tissue, harvested at different stages
during plant development.

Abstract

Sweetpotato (Ipomoea batatas (L.) Lam) ranks seventh in annual production among food crops in the
world. It is also an alternative source of industrial materials such as starch, ethanol and useful
components. Sweetpotao was recently reevaluated as one of the best healthy foods by the nonprofit
CSPI (2007) and the best bioethanol crop on marginal lands by USDA (2008), since it contains high levels of various
antioxidants such as vitamin C and carotenoids. Despite its importance as the well-being food source, little research has
been carried out on sweetpotato antioxidants at the molecular level. In this respect, transgenic sweetpotato with high
yields of ß-carotene by down-regulation of β-carotene hydroxylase (CHY-β) and lycopene ε-cyclase (LCY-ε) were
successfully generaed. We have recently isolated IbOrange gene responsible for carotenoid accumulation from the
orange-fleshed sweetpotato and introduced it into purple-fleshed sweetpotato to produce both anthocyanin and
carotenoids in one storage root. In the presentation, our recent results on metabolic engineering of pigment antioxidants
in transgenic sweetpotato will be introduced in terms of sustainable development on global marginal lands.

Abstract

Soil bacteria Sinorhizobium meliloti had enormous agricultural value,due to their ability in
fixing nitrogen symbiotically with an important forage crop legume- alfalfa.The aim of this
study (i)isolate indigenous S.meliloti from different field sites in Iraq,(ii) evaluate the isolates
tolerance to induced drought using polyethylene glycol-6000, (iii)assessing genetic diversity
and genetic relationships among isolates of natural population with drought tolerant abilities. Drought tolerance
study revealed vast variations between Sinorhizobium isolates, the highest tolerant isolates to drought were twelve
from total thirty (40%),tolerated from -3 up to -4 Mpa(mega pascal),while the drought sensitive isolates tolerated
up to – 1.5 Mpa,except isolate Bs58 which tolerated up to -1 Mpa water potential.The growth declined with the
increase of drought stress. Cluster analysis based on RAPD-PCR showed significant differences among S. meliloti
isolates, and the results gave almost identical grouping of isolates in regards to drought experiment. Among
indigenous isolates two divergent groups could be determined,the first major group included drought tolerant
isolates and the second major group comprised all drought moderate and sensitive isolates with 40% similarity
between the two major groups.

CONFERENCE ABSTRACT (SESSION)

Plant and Environment

THE ROLE OF CELLULOSE-SYNTHASE-LIKE CSLF6 IN 1,3;1,4-β-D-GLUCAN
BIOSYNTHESIS AND OVEREXPRESSION IN WHEAT GRAIN TO INCREASE SOLUBLE
DIETARY FIBRE LEVELS.

S.A. Jobling

CSIRO Agriculture Flagship, Canberra Australia

Abstract

Dietary fibre is largely derived from the cell walls of plants and in cereal grains, arabinoxylan and
1,3;1,4-β-D-glucan (betaglucan) are the major cell wall polysaccharides. Dietary fibre is an essential
part of a healthy diet however fibre consumption in most Western countries is below target levels.
Food products made from grains that have a high content of water soluble betaglucan such as oats
and barley are allowed specific health claims in some countries related to the lowering of blood
cholesterol. Wheat grain has only low levels of betaglucan, most of which is insoluble. The cellulose-synthase-like
CslF6 gene is a major component of the betaglucan synthase and we will describe work to characterise the function of
this protein from various cereals in Nicotiana benthamiana leaf expression system. Each CslF6 gene produces a
betaglucan with a distinct structure and this affects water solubility. Overexpression of selected CslF6 genes in wheat
grain increases levels of soluble betaglucan and dietary fibre.

Abstract

Mulgeum intake station located in downstream the Nakdong River supplies raw drinking water of
Busan metropolitan city in South Korea. As eight weirs were constructed by ‘Four Major River
Restoration Project’ in Nakdong River during 2009~2012, water environment of Nakdong River had
changed significantly. Therefore, the changes of algae occurrence and water qualities were monitored
and analyzed to evaluate the effects of weir construction on water environment using the each three years’ data before
and after of weir construction. As a result, water qualities in downstream the Nakdong River were improved after weir
construction. Chl-a, BOD, T-P and PO4-P concentration were significantly decreased after weir construction. Especially
the average Chl-a concentration was decreased from 119 mg/m3 to 47 mg/m3 in winter. This phenomenon of Chl-a
decrease is well corresponded with the decline of T-P and PO4-P. And it is considered that the improvement of water
qualities in downstream Nakdong River is affected with the cut off effect of nutrients by the weirs in midstream and
upstream. So algae occurrence in upstream and midstream the Nakdong River should be studied later.

CONFERENCE ABSTRACT (SESSION)

Plant and Environment

A WHOLE PLANT APPROACH FOR NITROGEN-FIXATION IN NON-LEGUMINOUS CROPS

Munusamy Madhaiyan and Lianghui Ji

Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore

Abstract

Jatropha curcas is promising new non-food crop for biodiesel production because of its ability to
thrive on marginal land, with strong tolerance to draught and poor soil nutrient content [1-3].
Nevertheless, high fertilizer input remains essential in order to have commercially acceptable oil
productivity. This reduces the green index of Jatropha biofuel. To address this issue, we studied the
taxonomical distribution of 1017 cultivable endophytic bacterial strains isolated from different parts of Jatropha with an
emphasis on nitrogen-fixing bacteria. Like other reports, we found strong tissue preferences of the bacteria in Jatropha.
The16S rRNA gene sequences can be assigned to five major phyla and, surprisingly, 31.2% of them potentially
represent new taxa. Nitrogen fixing isolates were found diverse and present in five classes belonging to α, β, γ-
Proteobacteria, Actinobacteria and Firmicutes. The phylum Proteobacteria was the most dominant amongst strains that
were positive for both nifH gene and endoglucanase activity. Methylobacterium species account for 69.1% of the leaf
endophytic bacterial isolates. Notably, many Methylobacterium isolates were able to fix nitrogen.

We will present genomic, physiology and plant-bacteria interaction studies on two strong nitrogen-fixing isolates:
Kosakonia sp. R4-368 (previously Enterobacter R4-368 [4,5]) that mainly colonizes in roots and stems and Methylobacterium radiotolerans L2-4 that mainly colonizes leaf tissues both as endophyte and epiphyte. Root treatment
of R4-368 or foliar application of L2-4 significantly improved growth parameters, such as plant height, leaf number,
relative chlorophyll content and stem volume. Importantly, strain L2-4 improved seed yield by 222.2% and 96.3% in
plants potted in sterilized and non-sterilized soil pots respectively. Strain R4-368 improved seed set by approximately
177% and 49.0% in sterilized and non-sterilized soil respectively. The average single seed weight was increased
approximately 10% by strain R4-368. Yield improvements were mainly attributed to an increase of female-male flower
ratio, which led to a corresponding increase of fruit and seed sets. Furthermore, there was an additive effect for seed
yield of root and leaf treatments were both performed. We will present the effects of application of the two isolates on
other crops.

CONFERENCE ABSTRACT (SESSION)

Plant and Environment

THE DECONSTRUCTION OF SEED MUCILAGES

Rachel A. Burton

Australian Research Council Centre of Excellence in Plant Cell Walls, School of Agriculture, Food and
Wine, University of Adelaide, Urrbrae, SA 5064, Australia

Abstract

Some plant seeds produce mucilage upon contact with water. Two well-known examples are
Plantago ovata and Chia (Salvia hispanica). The dry mucilage from Plantago is removed from seeds
by milling and marketed as Metamucil® whilst Chia is sweeping across the planet as a “superfood”;
both are promoted as being beneficial for human health, including for the control of Type II diabetes,
coronary heart disease and colon cancer. These claims are based on the bacterial fermentation of the polysaccharides
within the mucilage, once it reaches the large intestine. We have undertaken a detailed biochemical analysis of both
mucilage types and have carried out in vitro fermentation studies, allowing metagenomic profiling of the bacterial
species able to metabolise the polysaccharide components. Transcriptome analysis is also underway to identify genes
encoding polysaccharide synthases, information also relevant to economically important cereals wheat and barley which
contain similar polysaccharides in the grain. Genes from Chia may be particularly interesting as this mucilage contains
novel polysaccharide components that haven’t been identified in other higher plants.

Abstract

Degradation of poorly water soluble hydrocarbons, like n-alkanes and polycyclic aromatic
hydrocarbons are challenged by some bacteria through emulsification of hydrocarbons by bacterial
bio surfactants. In diesel oil bioremediation, diesel oil degrading and surfactant producing bacteria are
used to eliminate these pollutants from contaminated waters. Therefore, identifying and
characterizing bacteria capable of producing surfactant and degrading diesel oil are pivotal. In this study, bacteria
isolated from hydrocarbon contaminated river water were screened for their potential to degrade diesel oil. Primary
selection was carried out by using conventional enrichment culture technique, emulsification index measurement,
gravimetric and gas chromatographic (GC) analyses of diesel oil degradation. A bacterium with 60% emulsification
index and 92% diesel oil degradation ability in 14 days was identified as Acinetobacter haemolyticus Zn01 by 16S
rRNA sequencing. A. haemolyticus Zn01 was shown to harbor both catabolic genes alkb and C23O effective in diesel
oil degradation. The bio surfactant of the bacterium was also characterized in terms of surface tension, zeta potential,
fourier transform infrared spectroscopy and scanning electron microscopy. Being able to degrade diesel oil with a high
emulsifying index A. haemolyticus Zn01 seems to have high potential for the elimination of diesel oil from polluted
waters.

Plant and Environment (Track)

LONG TERM EVALUATION OF ALGAE OCCURRENCE IN DOWNSTREAM THE NAKDONG
RIVER, SOUTH KOREA: 1998~2014

Civil and Environmental Engineering, Pusan National University, South Korea

Abstract

Nakdong River is one of the major rivers in South Korea. Mulgeum site, which is downstream
of Nakdong River, is the intake station of Busan metropolitan city. This study examined the
chlorophyll-a(chl-a) concentration as a representative parameter of the occurrence of algae at
the Mulgeum site from 1998 to 2014. The water quality, hydraulics and climate conditions were
used to determine the parameters significantly affecting the chl-a concentration. The correlations between chl-a
and the other parameters were analyzed by the Pearson coefficient. As a result, the chl-a concentration showed
clear patterns according to the seasonal changes. High chl-a concentrations by Stephanodiscus spp., which is the
dominant species in low water temperature at Mulgeum site, were observed in winter. On the other hand, low chla
concentrations were observed in summer. The Monsoon climate characteristics have a significant effect in
summer. Correlation analysis also showed different significant parameters to seasonal changes. In winter, pH, DO,
BOD, COD, and PO4-P showed a significant correlation with the chl-a concentration.

CONFERENCE ABSTRACT (POSTER)

Plant and Environment (Track)

TRANSCRIPTOME ANALYSIS OF WHEAT GLUME UNDER DROUGHT STRESS

Changxin Liu and Jinyin Lv

College of Life Sciences, Northwest A&F University, China

Abstract

The non-leaf organs of wheat performed higher tolerance than leaf in the adverse environment.
To investigate the drought tolerance in wheat spike, we investigated variations of gene
expression in the early filing period at 6d post anthesis (DPA) by using Affymetrix Wheat
Genome Array. 30, 768 probe sets of total 61,703 ones were identified of wheat glumes at the
mRNA level. 256 differentially expressed probe sets were identified. A total of 229 probe sets
were up-regulated (FC≥2.0) and 27 were down-regulated(FC≤0.5) under drought stresses. Most up-regulated
genes were involved in functions such as signal transduction, metabolism and transcription. Among the upregulated
genes, we identified signaling proteins, transcription factors and abiotic stress-related genes. The signal
pathway networks constructed with KEGG showed three important genes involved in the phenylalanine
metabolism, in which phenylalanine ammonialyase, cinnamic acid hydroxylase, chalcone synthase and chalcone
isomerase located at the center of the pathway, which indicated their pivotal roles in the phenylalanine metabolism
and flavonoid biosynthesis derived from carbohydrate metabolism. Real-time PCR analysis confirmed that 10
genes were up-regulated and down-regulated at 0, 6, 12, 18, 24 DPA. Most up-regulated genes were involved
significantly regulated by drought stresses were of unknown function; our results provide new insights into the
molecular mechanism of the wheat spike responses against drought stress.

RESEARCHES REGARDING THE USE OF COACERVATES ESSENTIAL OILS IN SEED
TREATMENT IN ECOLOGICAL CROP PRODUCTION

Faculty of Agriculture, University of Agronomic Sciences and Veterinary Medicine, Bucharest,
Romania

Abstract

Following the continuous increase of population, the request for assuring the enough food is a
fundamental objective for all farmers. In these conditions is increased as well the request for
ecological food production. Answering to this problem, the objective of this research paper was to
establish if the microencapsulated essential oils, obtained by us in the experiments at laboratory scale,
may be used to protect the wheat seed from attak of pathogens and pests in soil after seeding. The research was
performed in laboratory and in experimental field.

The laboratory research was focused on microencapsulation of essential oils and on testing the phytotoxicity effect of
microcapsule upon the wheat seeds and as well for test upon the population of melolontha melolontha(MM) larvae stage
I, II. The microcapsules were obtained by a process of complex coacervation, and they have a central core formed by an
essential oil, covered with a shell made of crosslinked hydrolyzed collagen. In the experimental field was evaluated the
intensity of pests attack frequency. The results of phytotoxicity tests show that all the products used in seed treatments
don't have any phytotoxic influence at the used doses, on the contrary we identified a stimulating effect upon the plant
growth.

The researches regarding the influence upon the MM populations show the products with essential oil from thymus
vulgaris, satureja hortensis, Ocimum basilicum decreasing the intensity of attack by comparing with control variant
without any treatment. By comparing with control treated using imidacloprid, in laboratory, the efficacy of treatment
was around 50% from that one.

In the experimental field we found the decreasing significantly of dipterus attack frequency. Analyzing the results
obtained in this research we can said: "all the seed treatments based on own microencapsulated essential oils, can be
used successfully for wheat pest protect in ecological system"

REAL TIME PCR TO ELUCIDATE ENDOPHYTIC STAGE OF RAMULARIA COLLO-CYGNI
IN BARLEY SEED

Abstract

Ramularia collo-cygni (Sutton & Waller) (RCC) is a fungal pathogen infesting spring and winter
barley (Hordeum vulgare L.) and causes the disease known as Ramularia leaf spot (RLS) manifested
by necrotic spots on leaves. The symptoms usually appear during late growth phases of the host
plants. When severe, the disease progresses by very rapid development of spottiness. In addition to
reducing yields, it also decreases grain quality. A real-time PCR assay was designed to quantify RCC
in barley tissues. PCR primers and a TaqMan probe were designed to target an RCC-specific DNA sequence. The
method was optimized for pure fungal DNA and plasmid standard dilutions. Kernels were dissected into lemma,
pericarp, testa, endosperm and embryo, and a suspension was acquired after washing the kernels. All these parts were
tested for amounts of RCC, and the presence of other potential seed-borne pathogens was analyzed by standard PCR
diagnostics. The highest level of Ramularia DNA was in the lemma, and it occurred in lower amounts in the pericarp
and embryo. Ramularia DNA was also evidenced in the suspension after washing the kernels. Our findings indicate that
DNA of RCC is present on the emerging leaves of plants sown from infected seed. This confirms that RCC can be seedborne.
DNA of RCC is detectable by molecular methods also in cases when visible symptoms are absent. DNA content
in leaves need not necessarily correspond to the intensity of RLS symptoms. The work was supported by the Ministry of
Agriculture of the Czech Republic, Projects QJ1310091, QJ1310226, QJ1210008 and RO0211.

Abstract

The timing of floral transition determines successful reproduction in higher plants and is precisely
monitored by multiple environmental cues. The GIGANTEA (GI) gene participates in the network
connecting environmental stress and developmental stage transition in Arabidopsis. We first
identified GI gene orthologous from the perennial growth poplar (Populus alba × P. glandulosa) as
PagGIa, PagGIb and PagGIc. PagGIs are predominantly localized at nuclear and its transcripts are diurnally regulated
with a peak around zeitgeber time 12 under long-day conditions. Ectopic over-expression of the PagGIs in wild-type
Arabidopsis plants resulted in early flowering and enhanced salt sensitivity; while gi-2 mutant Arabidopsis plants
expressing PagGIs are rescued from delayed flowering and enhanced salt tolerance. Furthermore, PagGIs negatively
regulate the salinity stress tolerance by interacting with PagSOS2 in poplar. In addition, transgenic poplars with downregulation
of PagGIs by RNAi display enhanced salt tolerance. Taken together, our results reveal new insights on the
link between flowering time regulator and adaptation to salt stress in poplar.

CONFERENCE ABSTRACT (POSTER)

Plant and Environment (Track)

PHYLOGENETIC ANALYSIS AND MARKER DEVELOPMENT USING COMPLETE
CHLOROPLAST GENOME SEQUENCES OF SOLANACEOUS SPECIES

Tae-Ho Park and Kwang-Soo Cho

Department of Horticulture and Institute of Life and Environment, Daegu University, Gyeongsan
712-714, Republic of Korea

Abstract

Somatic hybridization has been widely applied to introgress several disease resistance from wild
Solanum species into the cultivated potatoes (Solanum tuberosum) and plastome genotyping is one of
important processes to select proper breeding lines. In this study, we completed chloroplast genome
sequences of two wild Solanum species, S. commersonii and S. nigrum, which have been known and
used as one of useful resources for improving resistance to several diseases. The sequences were compared with those of
Capsicum annum, Nicotiana tabacum, S. lycopersicum, S. bulbocastanum and S. tuberosum to develop InDel markers
for the application in cytoplasm genotyping. Although gene contents and their relative positions were almost same,
detailed comparison of their sequences identified several indels between S. commersonii / S. nigrum and S. tuberosum in
the intergenic and intragenic regions. Based on the sequence information of the indels, one and seven allele specific
RCR-based markers were developed for discrimination of S. commersonii / S. nigrum from S. tuberosum and the
markers were further confirmed with filial generations and other wild Solanum species, respectively. Additionally, the
phylogenetic tree revealed S. commersonii is located in a same node with S. tuberosum, but S. nigrum was slightly
further from the node.

Abstract

Carotenoids have essential roles in light-harvesting processes and protect the photosynthetic
machinery from photo-oxidative damage. We have previously isolated sweetpotato Orange gene
(IbOr), which confers accumulation of carotenoids and salt stress tolerance. Here, we reveal that IbOr
functions as holdase chaperone and involves in carotenoids accumulation by stabilization of
sweetpotato phytoene synthase (IbPSY), one of the key enzymes in the carotenoids biosynthesis pathway. The transcript
levels of IbOr gene were increased in stem, root and calli of sweetpotato after heat stress. IbOr contained DnaJ like
cysteine-rich zinc finger domain, was localized in both the nuclear and the chloroplasts, and interacted with IbPSY in
chloroplasts. Interestingly, IbOr was predominantly localized in chloroplasts after heat stress. The holdase chaperone
activity of IbOr enhanced the stability of IbPSY protein under the heat and oxidative stress conditions. Furthermore, the
transgenic Arabidopsis overexpressing IbOr exhibited increased tolerance to heat and oxidative stresses. These results
showed that holdase chaperone activity of IbOr plays a role in the regulation of IbPSY stability, carotenoids
accumulation and abiotic stress tolerances. Our findings thus suggest that IbOr will be contributed to develop plants with
enriched carotenoids contents and enhanced stress tolerance.

CONFERENCE ABSTRACT (POSTER)

Plant and Environment (Track)

ISOLATED AND FUNCTION ANALYSIS OF GAPC GENES AND THEIR PROMOTERS IN
DIFFERENT WHEATS

Fenling-Liu1, Min-Zhang, Shushen-Yang, Xiping-Deng and Lun-Shan

College of Life Sciences, Northwest A&F University, China

Abstract

In recent years, the cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC) have been
emerged as a multifunctional gene with defined functions in numerous subcellular processes,
especially in abiotic stress tolerance. But several of them also had been considered as a constitutive
housekeeping gene. We have previously identified six GAPC genes from wheat (Triticum aestivum
L.) and the result of qRT-PCR showed that four of the TaGAPC genes could be a reference gene. In
this study, to elucidate the regulatory mechanisms of similar reference gene in different wheats, two corresponding
promoters of TaGAPC genes were isolated, which were 87.54% identical. Different cis-acting elements were harbored
in two promoters, such as series dehydration and hormone responsive elements (MYBCORE、WREY and
ASF1MOTIFCAMV) in P1683 and cold induced response elements (MYC and LTRCORE) in P1693. To further
verification, the promoters were fused to GUS reporter gene and Agrobacterium-mediated transient expression assay
were conducted. The GUS staining showed that both the two promoters could drive GUS gene and tissues transiently
transformed by TaGAPC promoter-β-glucuronidase (GUS) gene fusion were differentially activated by NaCl, abscisic
acid and low temperature. The results suggested that promoters from different reference gene contributed different to
abiotic stresses.

Two weeks prior to the single 5 ½ hour counseling, a senior physician phoned each participant to establish disease
presentation, severity and establish rapport. Each was asked to bring a spouse or partner. Participants learned the
integrity of endothelial cells is essential for vascular health. They understood oils, meat, fish, fowl, dairy products, eggs,
nuts, avocado and sugary foods impair vascular protection. A plant-based nutrition expert reviewed food acquisition and
preparation. Each participant received a notebook with a copy of power point slides, peer reviewed studies, recipes and
strategies and a book with plant-based recipes. A lunch followed ending with question and answer session. Two prior
successful participants gave testimonials. Exercise was encouraged. We invited follow up communication.

Other Areas (Track)

TOWARDS A UNIVERSAL METHOD FOR PROTEIN REFOLDING

Abstract

Escherichia coli is one of the most widely used hosts for the production of recombinant proteins of
therapeutic or commercial interest dedicated to structural and functional analysis. However, this
expression system is often hampered by the formation of insoluble protein aggregates (inclusion
bodies). In vitro refolding of such proteins into their native states requires screening of numerous
experimental parameters specifically optimized for each system. Hence, there is currently no reliable straightforward and
universal experimental solution providing the optimal refolding of proteins. The development of new original techniques
in this field is therefore crucial.

In that context, we have successfully demonstrated the reliability of a new procedure for protein refolding [1-3]. This
peculiar protocol is based on the association of an ionic detergent with a cosolvent. Indeed, though being known to
feature denaturing abilities, some detergents appear to have their properties strongly altered when interacting with a
cosolvent, and strikingly an appropriate combination of both even turns to a refolding of the protein. This remarkable
procedure has successfully been applied to soluble α-helix and β-sheet peptides, as well as soluble and membrane
proteins with several types of structures and properties.

In this contribution, we summarize our progress in the understanding by experimental (spectroscopy) and theoretical
methods (molecular dynamics), of our recently reported approach.

Abstract

Tryptamine Schiff bases were prepared by the condensation reaction of ethanolic solution of
tryptamine with 5-chlorosalicylaldehyde, 5-nitosalicylaldehyde and 3,5 ditertiarybutyl salicylalehyde.
The ligands were coordinated to Cu(II), Ni(II), and Zn(II). All the ligands and complexes prepared
have been characterized by spectroscopic methods such as IR spectra, 1H and 13C NMR, UV-Vis,
mass spectra, elemental C H N analysis, X-ray crystallography and magnetic susceptibility
measurements, as well as atomic absorption spectroscopy for metal analysis. Spectroscopic data shows that the ligands
are bonded to Zn and Cu metal centers via N and O atoms in a distorted tetrahedral geometry, whereas square planar
geometry is proposed for Ni complexes.

The biological activities of the ligands and complexes as anti ulcer agents have been investigated using Cimetidine as a
standard drug. The screening was done on high and low doses (60mg/kg and 30mg/kg body weight respectively) on
Sprague-Dawley rats which had been ulcer-induced by absolute ethanol. The ulcer lesion of the stomachs were identified
and counted and then correlations between the effects of the different compound have been made. The results revealed
very effective inhibition of gastric ulcer for ligands and complexes.

CONFERENCE ABSTRACT (SESSION)

Other Areas

HELIO-GERO PROTECTED DRINK-WATER AND FOOD: THE NEW POSSIBILITIES AND
PERSPECTIVES OF THE PREVENTIVE MEDICINE

Alexander Trofimov

International Scientific Research Institute of Cosmic Anthropoecology (ISRICA), Novosibirsk,
Russia

Abstract

In recent years, in the era of nano-technologies of information and knowledge, in Novosibirsk
intensively conducted research biotropic weakly energy avant-garde influences on the physiological
condition of the plants, animals and humans heliogeophysical numerous factors. Scientists of ISRICA
has developed and patented the technology for creating holograms with physiologically active
information (Patent RU 2239860,2004,A.Trofimov,K.Evstratov, 2015) sing of a new type of hologram, presumably,
containing "analog information about the quantum states" of antioxidants, vitamins, amino acids and other essential
compounds of the best examples of the cluster structures of clean drinking water, provided the use of the original
screening device "TRODR"( autors A.Trofimov, G.Druzhinin,2011)[1] repeatedly weakening of the magnetic induction
field of the Earth, it was possible to create a technology to produce helio-hero tread water, which reduces the functional
dependence of biological systems, including humans, are very high, often pathological helio-dependent from extreme
impacts of solar and geomagnetic disturbances ( Patent RU2342149). At last years we are created else one new
technological complex «HELIOSTAR» for the exposition at hypogeomagnetic conditions not only drink water, but and
the water contained food (seeds, milk, honey, caviar of fish).We want propose for the realization the new scientificpractical
program «STOP-TIME». The main aim of this program is the decreasing of speed of human aging. It is the
new scientific reality now!According to the our data , received with the use of only one in the world hypogeomagnetic
installation, and about 70% of the population of Siberia can be attributed to helio-magnetically sensitive people (high
and medium), responding to sudden changes heliogeophysical medium high blood pressure, and often hypertensive
crisis with severe consequences (heart attacks, strokes, etc.)[2]. At many congresses in gerontology, held in St.
Petersburg, Amsterdam, Paris, in the first decade of the XXI century, were discussed the new scientific data these units
on the dependence of the human life-span of the solar activity in the period of intrauterine development. It is shown that
the «threshold of longevity reach, mainly those of intrauterine development which took place with minimal activity of
the Sun[3,4]. These data were obtained from the use of the developed and registered in Russia of the computer program
«Helios» (ISRICA, SCCEM RAMS ), the world community was first introduced to the world EXPO- 92 in Sevilla
(Spain)[5]. How do you change the «Sunny code», which prevents the solar-addicted people «get sick less and slower
aging»? Effective non-drug prevention helio-magnitotropic reactions and states rights, as well as possible ways to slow
the rate of aging of the body have not existed until quite recently .Now for this role claims helio-geroprotective water,
developed and tested in ISRICA.It is a real help in the confrontation, in this sense, a defenseless person, over the raging
solar element!This a protective tool can be indispensable to all of us, as the crew spacecraft «Earth» in collapsible
currently very dangerous for all living cosmic-physical situation, when the magnetic field of the Earth, this natural buffer
against solar corpuscular flows continues to decrease and, accordingly, more and more high-energy particle reaches the
upper layers of the atmosphere, causing showers of secondary ionized corpuscles, threatening the biosphere and man[6].
We have shown that the plants watered by helio-tread water, and also Wistar rats , who took this water for drink, unlike
the controls, show a high resistance to stress in animals males and we noticed increase testosterone in blood and tissues
and relative abundance of stable isotopes of carbon (13 C) which usually with age irreversibly lost by the body[4]. In
people with hypertension after 2-3 weeks of taking this water manifested distinct helio-tread effect: in periods of solar
and geomagnetic storms they have not reacted to the solar-magnetospheric disturbances rise in blood pressure and
deterioration of their health, so they went out of the zone of risk of development of crises and many complications [7].
Helio-trade effect was confirmed by rigorous mathematical analysis, when was observed significant INVERSE
correlation between the majority of the functional parameters of the man and intensity of cosmophysical factors. Exist
the necessity for the creation of a global system of geoecological human life support in the conditions of spreading
heliogeophysical changes of our biosphere on the basis of new biotechnologies.

Other Areas (Track)

DETERMINE THE ROLE AND MECHANISM OF GENE CODON USAGE IN REGULATING
PROTEIN EXPRESSION AND FOLDING

School of Biotechnology, East China University of Science and Technology, Shanghai, China

Abstract

During heterologous protein expression, the aim is to increase expression levels without altering
native activities. Gene expression is mostly known to be regulated at the transcriptional level, while
protein function can be regulated post-translationally. However, as a co-translational mechanism, the
role of codon usage in regulating protein expression and function is less well studied.

In many organisms, there is a bias for preferred codons and it is thought to be a mechanism to enhance expression for
highly expressed genes. For example, Neurospora crassa prefers to use G/C rather than A/T at the 3rd position of codons
in highly expressed genes. FREQUENCY (FRQ) is a key component in the negative feedback loop of Neurospora crassa
circadian clock. Compared with other genes, frequency (frq) exhibits a very non-optimal codon usage. It's not clear
whether this "poor" codon usage has any biological significance.

To test the role of codon usage in frq, here we made a series of codon optimized frq constructs, and introduced them into
a frq null strain. We found that strains carrying the N-terminal optimized frq exhibited severe circadian clock
phenotypes, with higher FRQ protein levels but similar frq mRNA levels compared with wild-type. Surprisingly, shown
by both trypsin sensitivity assay and freeze-thaw assay, FRQ proteins in these strains were less stable. Besides, they
were defective in binding their WCC partners, and optimized FRQ had a compromised function to support WC-1 and
WC-2 levels in the positive feedback loop, suggesting that codon optimization also influences protein function even
though its sequence is unchanged. Taken together, codon usage of frq regulates its protein expression, folding and
function. Therefore a scientific analysis is needed during codon optimization design when expressing heterologous
proteins.

Abstract

The synthesis, characterization, thermogravimetric and antioxidant activity studies of Schiff bases
derived from 4-phenyl-1,3,5-triazine-2,6-diamine and 3-hydroxy salicylaldehyde (H2L1), 4-hydroxy
salicylaldehyde (H2L2) together with their nickel (II) and zinc (II) complexes are reported. The
ligands were characterized by IR, 1H and 13C NMR, UV-Vis spectroscopy as well as elemental
analysis. Spectral studies reveal that the ligands were acting as tetradentate chelating agents and
coordinated to the metal center via deprotonated phenolate oxygen and azomethine nitrogen atoms in a 1:1 ligand to
metal ratio. The antioxidant activities of the ligands and complexes were examined using the DPPH radical scavenging
method. The results show that the ligands exhibit higher radical scavenging ability than the complexes.

DIMETHOXYFLAVONE, A FLAVONOID FROM STEREOSPERMUM KUNTHIANUM STEM
BARK WITH ANALGESIC AND ANTIINFLAMMATORY ACTIVITIES

Abstract

Introduction:Stereospermum kunthianum is reputed for its ethnomedicinal use in the treatment of
various painful and inflammatory conditions in Nigeria and other West African countries [1].

Materials and Methods: The chemical identification and isolation of the stem bark of
Stereospermum kunthianum led to the isolation and characterization of an alklylated flavonoid
(3,7,4/-trihydroxy-3/-(8//-acetoxy-7//-methyloctyl)-5,6-dimethoxyflavone. The structure was confirmed by 2-DNMR [2]
and other spectroscopic techniques [3].

The analgesic and anti-inflammatory activities of the isolated compound were studied using the Randall-Selitto test [4],
mouse writhing assay [5] and carrageenan-induced paw oedema in rats[6].

Results: Dimethoxyflavone at the dose of 25mg/kg and 50 mg/kg significantly increased (P < 0.001) the pain threshold
when compared with the rats that similarly had sham treatment with distilled water. The percentage increase in pain
threshold was 51.6, 66.7 and 62.3 respectively for dimethoxyflavone at 25 mg/kg, 50 mg/kg and indomethacin (10
mg/kg) respectively. In the mouse writhing assay, the writhes were significantly reduced (P < 0.001) in the
dimethoxyflavone treated mice with the percentage inhibition of pain of 46.2, 58.3 and 63.1 for the flavonoid at 25
mg/kg, 50 mg/kg and aspirin (100 mg/kg) respectively. Dimethoxyflavone (25mg/kg and 50 mg/kg) caused a significant
reduction (P <0.001) in the paw swelling as from the 2nd h and a much more reduction at the 3rd h post carrageenan
injection compared with control rats.

Conclusion: The result of the phytochemical study led to the isolation of 3,7,4/-trihydroxy-3/-(8//-acetoxy-7//-
methyloctyl)-5,6-dimethoxyflavone, a known flavonoid from Stereospermum. kunthianum stem bark which possesses
both analgesic and antiinflammatory activities.

MODULATION OF ENDOPROTEASES BY TRICHOPHYTON RUBRUM GROWTH ON
ELASTIN AND KERATIN SUBSTRATES

Abstract

Dermatophytes are keratinolytic fungi, classified as geophilic, anthropophilic and zoophilic species
on the basis of their primary habitat association. The anthropophilic specie Trichophyton rubrum is
the most common in superficial mycoses worldwide. Although dermatophytes do not normally
penetrate beyond the epidermis, deeper penetration and systemic infections can occur in
immunocompromised hosts. The epidermis of skin is composed by keratin, while derme is
constituted by collagen, fibers and elastin. The secretion of proteases is considered a key factor for virulence of
dermatophytes, and arthroconidia play a central role in pathogenesis, since is responsible for skin adherence. The aim of
this work was to verify the modulation of Subtilisin3 and Subtilisin1 in germinating conidia of T. rubrum growth on
elastin and keratin substrates. The modulation of two encoding genes of Subtilisin3 and Subtilisin1 was carried out by
qPCR using a pool of RNA of 24h, 36h and 72h of T. rubrum strain CBS118892 growth on keratin and elastin. The
results show that the Subtilisin3 is similar induced by T. rubrum in both protein substrates, whereas subtilisin1 is more
induced on keratin substrate. The modulation of these genes may be important to elucidate the superficial and deep
infection mechanism.

Abstract

BACKGROUND

In 2002, the author presented and subsequently published that with space flight endothelial
dysfunction; there is the potential for congestive heart failure despite invariable dehydration. There is
decreased thirst, inappropriate diuresis, atrophy of water storage sites in skeletal muscles and reduced plasma volume;
this may be triggered by endothelial intercellular gaps with inflammation in post -capillary venules. These leaks may be
responsible for an invariable 10% loss of plasma volume in microgravity. Furthermore, the invariable significant
reductions of the serum magnesium (p<0.0001) in large groups of astronauts and cosmonauts, despite very poor serum
sensitivity, along with catecholamine elevations to levels twice those in the supine position on Earth with ischemia and
multiple vicious cycles, are conducive to further endothelial injuries and in turn, to catecholamine cardiomyopathy
( acute temporary heart failure.)

SYNDROME

During his lunar last 20 minutes, Neil Armstrong notified Houston twice at 4 minute intervals that he was “short of
breath” at 111 hours and 32 minutes and “still short of breath” at 111: 36. This symptom occurred prior to the potential
confounder of inhalation of highly toxic iron-laden dust, brought into the habitat on space suits. Whereas his heart rate
on the moon was 130-160/ minute, just prior to splashdown in the Pacific, after his 3 day journey back to Earth, his heart
rate was down to 61. The explanation for this correction of tachycardia, could only be that by quenching his severe
thirst, the very high adrenaline levels were reduced because of expansion of the left ventricle by replenished plasma
volume with in turn reduction in the gradient; this could have been precipitated by protrusion of the septum into the left
ventricle as postulated by Merli et al.

CONCLUSION

Neil Armstrong Space Syndrome: Armstrong developed severe shortness of breath and very rapid heart rate on the
moon BEFORE inhalation of toxic iron-laden dust, brought into the habitat on the space suit.

1. SEVERE DYSPNEA, manifested by Armstrong’s notifying Houston twice during a 4 minute interval that he was
short of breath, consistent with acute congestive heart failure.

2. SEVERE THIRST; with microgravity, there is invariably decreased thirst, but in Armstrong’s case, despite the 3 day
trip back to Earth in microgravity, he apparently quenched his severe thirst.

3. SEVERE TACHYCARDIA CORRECTED BY FLUID REPLENISHMENT; with decreased plasma volume and in
turn increased adrenaline with tachycardia, restoring his plasma volume, would in turn, markedly reduce a high heart
rate.

CONFERENCE ABSTRACT (INVITED)

Regenerative Medicine

PERSONALIZED HEALTHCARE: FROM RESEARCH TO REALITY

Jeffrey P. Harrison and Debra A Harrison

Department of Public Health, Brooks College of Health, University of North Florida, 1 UNF Drive,
Jacksonville, FL 32224-2645, USA

Abstract

Over the past decade, healthcare researchers have spent hundreds of millions of dollars in sequencing
the first human genome. In addition, there is significant research in the use of stem cells to
regenerate specialized tissue for treatment of individual patients. This session will address the use of
this research to bring “Personalize Healthcare” to the patient. Specifically, we will discuss the use of
Human Genomics and other stem cell therapies to regenerate organs and treat pressure ulcers. The appropriate use of
these new clinical technologies provides opportunities for significant improvements in healthcare quality as well as
improvements in preventive health services. The growth of “Personalized Healthcare” requires the creation of
collaborative relationships between researchers, clinicians and healthcare leadership as process-focused care continues to
develop. The growth of “Personalized Care” facilitates innovation across the continuum of healthcare practice and may
be a cost effective approach to improving healthcare quality.

Abstract

Introduction: Thymus is important organ of the immune system. Fromdiscovery of thymosine alpha 1 play essential role in maturation of peripheral T lymphocytes. Manyfactorswasdiscoveredthat cause precocious thymus atrophy and developing of autoimmunediseases. Manymethods are described to stop autoimmunediseasesincludingrevolution in oncology ((Ipilimumab),and stem celltherapy.

This paperdescribe a new method to improvenatural thymus defence system by ectopic thymus derived stem cell implantation.

Material and Methods: Thymus explants werecultured for 34 days to observe secretion of thymosine alpha 1 in vitro and after implantation intovolunteers. Cellswereimplantedinto the subcutaneous adipose tissue and Tha1 levelwasobservedduring 12 years. Number of implantedcellswasadequate to the size of 3,0 thymus remnants of adult man.

Conclusion: Thymus progenitor/stem cellsmaybeused in future as a new additionaltoolls in case of thymus deficiency and in autoimmunediseases. Instead of veryexpensive pharmaco-therapyusingsyntheticthymosine (Zadaxin) and otherphatmaceuticals.

CONFERENCE ABSTRACT (SESSION)

Regenerative Medicine

NUMERICAL MODELING OF MECHANOTACTIC INFLUENCE ON CELL MORPHOLOGY

Mohamed Hamdy Doweidar, Seyed Jamaleddin Mousavi

Group of Structural Mechanics and Materials Modelling (GEMM), Aragón Institute of Engineering Research (I3A), University of Zaragoza, Spain.
Mechanical Engineering Department, School of Engineering and Architecture (EINA), University of Zaragoza, Spain.

Abstract

Cell morphology is a key aspect in many biological processes such as morphogenesis, tumor growth and wound healing. Among other cues, mechanical characteristic of the surrounding micro-environment can control the cell morphology. It is well known that traction forces transmitted to the extracellular matrix (ECM) through cell focal adhesions and integrins play a fundamental role in this process by rearranging the cell cytoskeleton (CSK). In this work we have developed a novel 3D computational model to comprehensively predict the evolution of cell morphology during migration due to mechanotaxis.

A discrete methodology is here chosen by which the cell is represented by a group of finite elements. Therefore, during migration, the cell shape can be efficiently remodeled in a free mode. The present model is developed based on equilibrium of the effective forces over the cell body; the traction force, the protrusion force and the drag force. The cell traction force is governed by the cell internal deformation. The random protrusion force is generated by actin polymerization. The drag force is the substrate viscous resistance.

Correlated with experimental observations, the present model illustrates that the morphology of an adherent cell can be controlled by substrate stiffness and boundary conditions. Our findings indicate that within an unconstrained substrate with a soft (several kPa) and hard (>200 kPa) stiffnesses, the cell is unable to adhere or penetrate into the substrate so that the cell remains mainly rounded without any specific preference of migration direction. In contrast, when a cell is located within a substrate with an intermediate (10 kPa) and rigid (100 kPa) stiffnesses the cell can actively adhere to the substrate migrating towards the constrained surfaces. It can be concluded that in the intermediate and rigid substrates the higher the traction force, the greater the cell elongation, the larger the cell membrane area, and the less random the cell alignment.

Acknowledgements: The authors gratefully acknowledge financial support from the Spanish Ministry of Economy and Competitiveness (MINECO MAT2013-46467-C4-3-R) and the CIBER-BBN initiative. CIBER-BBN is financed by the Instituto de SaludCarlos III with assistance from the European Regional Development Fund.

Abstract

One of the actually problems of tissue engineering and regenerative medicine is the search for
biocompatible materials having the desired properties. The goal of the conducted studies was proof of
the effectiveness of injectable biopolymer microheterogeneous hydrogels (IBMH): an independent
implantable system to replace soft tissue defects including the stimulation of the regenerative
processes of patient's own tissues; as a delivery system for the transplantation of cells and a temporary scaffold and a
nutritive media while creating the tissue-engineered constructs. Compositions of IBMH were obtained from the
hydrolysate of embryonic or postnatal tissues of animal using technology of ultra-dispersion hydrogels with subsequent
radiation crosslinking. The IBMH contains practically all high- and low molecular weight components of extracellular
matrix, namely, partially hydrolyzed collagen peptides, proteoglycans, and glycoproteins By varying the composition
and size of the microparticles of crosslinked hydrolysate from 30 μm to 300 μm, and the ratio of solid and liquid phases,
was established linear range of the IBMH with different rheological properties and time of bioresorption (from several
weeks to several months). Using AFM analysis of microparticles IBMH the porous structure of particles with a pore size
of 2-4 μm was found, which is a positive properties in the processes of neovascularization and neoinnervation into the
tissue-engineered constructs based on the IBMH. The results of experimental studies on the creation of cell-engineering
constructs of cartilage tissue, liver and pancreas reveal about the ability of the IBMH long time to support the activity of
the cells, including the processes of proliferation, differentiation, and synthesis of own ECM, which gradually replaces
by resorbed ECM mimetic.

CONFERENCE ABSTRACT (SESSION)

Regenerative Medicine

SCALE-UP OF IMMUNE CELL THERAPIES FOR ORGAN TRANSPLANTS

Marianne Ellis, Fadi Issa and Jon Pleat

Department of Chemical Engineering and Centre for Regenerative Medicine, University of Bath,
Bath, UK

Abstract

Organ transplants require the recipient to take immunosuppressant drugs for their entire life to
prevent rejection of the allogeneic tissues [1], which may cause side effects. Recently, work has been
carried out to harness the inherent immune control mechanisms [2] and an alternative to
immunosuppressant drugs is the administration of a large dose of natural regulatory T cells (Tregs)
[2]. A therapeutic number of cells (suggested to be 30 x 106 cells/kg [3]) need to be made available at the right time. The
current in vitro expansion process for Tregs is well established at bench scale using static multi-well plate culture; the
cells are split after 4 days then every day, cultured in the presence of Life TechnologiesTM Dynabeads®. A robust and
cost effective Treg expansion bioprocess would enable Treg therapy to replace immunosupressant drug therapy and so
improve the post-transplant outcome for the patient’s quality if life, reduce or eliminate additional NHS care due to
complications, and reduce the cost of transplantation. Here we present a semi-automated cost-effective process for Treg
cell therapy manufacture, based around a fluidized bed bioreactor.

IN VITRO RESTORATION OF SMN PROTEIN BY HOMOLOGOUS RECOMBINATION
METHOD

Abstract

The reduced level of Survival motor neuron (SMN) protein causes a neurodegenerative disorder
named as Spinal muscular atrophy (SMA) that characterized by progressive paralysis and
symmetrical muscle weakness. Majority of SMA patients have homozygous deletions in SMN gene.
Despite extensive efforts to find a cure for SMA, there is still no effective treatment for this
devastating disease. In the present study, 'gene targeting' method based on homologous recombination
was used to restoration of SMN protein expression in SMA type I fibroblasts. Gene targeting fragment including SMN1
cDNA and homolog upstream and downstream sequences was designed and subcloned into a pGH vector. SMA type I
fibroblast cell line (Coriel institue, GM03813) was transfected with DNA fragments using Lipofectamine LTX Plus
Reagent (invitrogen, 15338). Single cell colony was achieved by serial dilution and PCR analysis was performed to
confirm the occurrence of homologous recombination in transfected cells. Restoration of SMN protein expression was
confirmed by RT-PCR and Western Blot analysis. The immunofluorescent results demonstrated localization of SMN
protein in the nucleus of transfected cells. Our result show that homologous recombination could present as an
alternative method for in vitro restoration of SMN protein. In vitro correction of patients stem cell by homologous
recombination and transplantation of these cells to patients could be investigated to find alternative treatment for SMA
disease.

CONFERENCE ABSTRACT (POSTER)

Regenerative Medicine (Track)

Laboratory for Stem Cells, Croatian Institute for Brain Research, University of Zagreb, Croatia

Abstract

To reach a satisfactory level of survival and healing properties, it is hypothesized that transplanted
neural stem cells (NSCs) need to enter the process of formation of synaptic contacts with both host
neurons and in-between themselves. Thus we induced ischemic brain injury by middle cerebral artery
occlusion in C57/Bl6 mice, labeled cells with a fluorescent dye and stereotactically injected them 5
days after injury at the dorsal cortico-callosal boundary. Animals were sacrificed at 2, 4, 8 and 14 weeks following
transplantation and we analyzed expression of Cell adhesion molecule 1 (CADM1), Neuroligin 1 (NLGN1) and
Synapsin I (SYN1). Migration of transplanted NSCs from the implantation site to the site of the lesion was observed and
cells aggregated robustly in the ischemic core. They were positive for Nestin, and majority of them for CADM1 and
Neuroligin 1. No SYN1 staining was present, even after 14 weeks. This in vivo observed pattern was similar to our in
vitro results. NSCs are exhibiting a marked and early visible potential for interaction with other cells and extracellular
matrix which suggests that formation of synapses contributes to survival, integration and healing properties of NSCs.

ESTABLISHMENT OF A NOVEL TUMOR ENDOTHELIAL CELL LINE DERIVED FROM
HUMAN HCC

Abstract

Tumor angiogenesis plays a crucial role in cancer growth, recurrence and metastasis. However, the
studies on the blood vessel formation and regulation in tumor have been hampered by the lack of a
steady source of tumor endothelial cell (TEC). In our pilot study, we freshly isolated CD31+
endothelial cells from the tissues of human hepatocellular carcinoma (HCC) and cultured them in
ECM medium with several supplements, and found that TECs in primary culture proliferated to confluency in 15 days to
20 days and stopped growing when subcultured to passage 7. To establish a novel TEC line, the cultured TECs were
transfected with SV40-LT. The cloned TECs containing SV40-LT have been subcultured to passage 53 to date. The
novel cell line of tumor endothelial cell derived from human HCC, with a steady growing rate in ECM medium has been
established. These transfected TECs exhibited similar endothelial characteristics as the primary TECs, i.e. that appeared
the cobblestone and spindle shape, Weibel-Palade bodies in the cells, both CD31 and CD105 expression on the cell
surface, chromosome number with 44. Furthermore, functional assays showed that the tube formation and migration also
occurred in the transfected cells. These results suggest that a novel TEC cell line established here maintains endothelial
characteristics and potencies. Therefore, our TEC cell line should be useful for studying of tumor angiogenesis and
vascular targeting therapy.

Acknowledgment: This study is granted by the Natural Science Foundation of China (NSFC), Grant No. 81272378.

PAOT TECHNOLOGY® NEW APPROACH FOR DETERMINATION OF OXIDATIVE STRESS
STATUS OF BIOLOGICAL LIQUIDS AND TISSUES

Abstract

Are you "rusty"?

A unique technology in the world, PAOT skin Technology® developed by the European Institute of
antioxidants (IEA) - this is a world first - a device that allows to monitor the status of your cells and
the famous "oxidative stress "which would be responsible for many cardiovascular diseases, and
promote the development of cancer. The PAOT skin Technology® is able to measure the full concentration of oxidants,
the famous "free radicals", and antioxidants in biological tissues such as skin, plants. These valuable measures will allow
everyone to get an idea of his general condition, and if necessary change its lifestyle, playing on well-known parameters:
physical activity, smoking, alcohol, stress, diet ... and eventually to better control the use of emulsions and various
creams available on the market, and consume reasonably tea, fruit juice, coffee, and other dietary supplements.

Various tests used to assess the antioxidant capacity but they are based on different mechanisms using different sources
of radicals or oxidants thereby generating different values not directly comparable. Indeed, these tests do not take
account of the diversity of antioxidants (including bioavailability and biological effects are varied), nor the hydrophilic
or hydrophobic nature or the specificity of interaction has alleged targets.

The work describes a PAOT skin Technology® for determining oxidative stress statute of biologicals solutions and
tissues.

Abstract

Angiotensin-I-converting enzyme (ACE) plays a key role in blood pressure regulation. It involves in
the conversion of angiotensin-I to potent vasoconstrictor (angiotensin-II) as well as inactivates
vasodilator (bradykinin). For the remedy of hypertension, ACE inhibitory drugs have been used. This
project is designed to isolate and purify ACE inhibitors from caprine milk which are important for the
treatment of hypertension. Caprine milk protein (CMP) was hydrolysed to release ACE inhibitory peptides (ACE-IP) by
employing different proteolytic enzymes i.e trypsin, pepsin and both in combination at different time intervals. Pepsin
showed highest ACE inhibitory activity 87.84% after 36 hrs. By using gel filtration chromatography (sephadex G-25)
ACE-IP were purified and got eight fractions that exhibited ACE inhibitory activity. The Fraction number 64 from CMP
hydrolysates by pepsin (36 hr) was showed the maximum ACE-inhibition activity. RP-HPLC was used to analyze the
purified ACE-IP. One of the most active fractions in relation to protein content was F64 with IC50 8.7 mg/mL. The
sequences of amino acid were also determined by amino acid analyzer.

CONFERENCE ABSTRACT (IN PROCESS)

Other Areas (Track)

PHARMACOGENOMICS OPEN A NEW GATE WAY IN THE TREATMENT OF CELIAC
DISEASE

Somia Gul, Abeeha Malick, Sameera Kaleem and Aniqa Malick

Faculty of Pharmacy, Jinnah University for Women, Pakistan

Abstract

Background: An immune mediated enteropathy celiac disease (CD) is an auto immune deficiency
disease that is occurs due to damage to the lining of the small intestine when foods with gluten are
eaten. Celiac disease is characterized by musculoskeletal problems digestive problems, a severe skin
rash called dermatitis, herpetiformis, iron deficiency anemia (low blood count), growth problems and
failure to thrive (in children), Seizures, Aphthous ulcers, tingling sensation in the legs.
Objective: Our goal is to emphasize on the treatment confusion due to misjudgment with celiac disease and irritable
bowel syndrome and its treatment that become possible not only by the use of gluten free diet but can also be treated by
emerging field of pharmacogenomics technology.
Methodology: In our survey we communicated with Pakistan celiac society, we also negotiated with Abbasi Shaheed
Hospital gastroenteritis specialists who are working on this disease for past year. We analysis the general and behavioral
or central nervous system conditions as well.
Result: The result shows that CD is misdiagnosed with IBS 42% consider it a small intestine disease while 58 % thinks
it's a large intestine disorder. 73% say diagnosis can be done via antitransglutunabase anti body, 88% says TGG, igA can
be helpful in detecting Celiac disorder, 48% favors endoscopy while biopsy is recommended by 52%.The outcome of
this innovative research is that celiac disease that occurs genetically influence in people that can be defecate by the
vaccination and people enjoy their life without any restriction.
Conclusion: The prevalence study is enormous research for celiac disease. There is no study yet on national or
provincial scale that reflects any data about the number of patients existing in Pakistan. The pharmacogenomics
advancement opens a new hope for the celiac disease patients, therapeutic vaccine that contains three proprietary
peptides that elicit an immune response in patients with celiac disease who carry the immune recognition gene HLADQ2.
Due to lack of awareness and varying symptoms of celiac disease make it hard to diagnose and creating a problem
in timely diagnosis and management. Currently it is leading to many problems including proper diagnosis and post
diagnosis management.

ACHIEVEMENTS AND CHALLENGES IN DEVELOPMENT OF WILDLIFE FORENSICS IN
SOUTH-EAST ASIA FOR CONTROLLING ILLEGAL TRADE FOR BIODIVERSITY
CONSERVATION: A CASE STUDY FROM INDIA

Abstract

Functional food is a natural or processed food contains known biologically active compounds, which
provides a clinically proven and documented health benefit, and thus, an important source in the
prevention, management and treatment of chronic diseases of the modern age. In this experiment
Spirulina, a well known functional food worldwide is irradiated by 60Co gamma radiation by
different doses 5, 10, 15, 20, 25 and 30 kGy at a dose rate of 5 kGy per hour. Amongst the irradiated samples Spirulina
exposed to 15kGy showed most inspiring results both in vitro and in vivo. 15kGy irradiated Spirulina showed maximum
fat binding capacity (FBC,47%) and sugar binding capacity (SBC,119%). Water holding capacity, water retention
capacity and swelling capacity of 15kGy irradiated Spirulina was found to be highest and were 33.68%, 40.22% and
86.96% respectively. Mice fed with irradiated Spirulina, exhibited lower weight gain, reduced blood glucose, TG,
cholesterol, LDL level and higher HDL level compared to the control group although both of the groups were fed fat
rich diet.

CONFERENCE ABSTRACT (IN PROCESS)

Other Areas (Track)

BOOSTING THE FOOD FUNCTIONALITY (IN VIVO AND IN VITRO) OF SPIRULINA GROWN
IN BANGLADESH BY GAMMA RADIATION: AN INSPIRING APPROACH

Hossain Uddin Shekhar, Emran Kabir Chowdhury and Jahid M M Islam

Biochemistry and Molecular Biology, Dhaka University, Bangladesh

Abstract

Functional food is a natural or processed food contains known biologically active compounds, which
provides a clinically proven and documented health benefit, and thus, an important source in the
prevention, management and treatment of chronic diseases of the modern age. In this experiment
Spirulina, a well known functional food worldwide is irradiated by 60Co gamma radiation by
different doses 5, 10, 15, 20, 25 and 30 kGy at a dose rate of 5 kGy per hour. Amongst the irradiated samples Spirulina
exposed to 15kGy showed most inspiring results both in vitro and in vivo. 15kGy irradiated Spirulina showed maximum
fat binding capacity (FBC,47%) and sugar binding capacity (SBC,119%). Water holding capacity, water retention
capacity and swelling capacity of 15kGy irradiated Spirulina was found to be highest and were 33.68%, 40.22% and
86.96% respectively. Mice fed with irradiated Spirulina, exhibited lower weight gain, reduced blood glucose, TG,
cholesterol, LDL level and higher HDL level compared to the control group although both of the groups were fed fat
rich diet.

CONFERENCE ABSTRACT (IN PROCESS)

Other Areas (Track)

College of Chemical and Biological Engineering, Zhejiang University, Zhejiang, China

Abstract

Volatile organic compounds (VOCs) and odorous compounds discharged into the environment create
ecological and health hazards. Among VOCs, a considerable number of BETXs (benzene,
ethylbenzene, toluene and xylenes) into the environment originates from industrial facilities. Organic
odorous pollutants are usually related to mercaptans (R-SH), organic sulfides (R-S-R), and disulfides
(R-S-S-R). Due to the high volatility, potential toxicity and the extensive use of these organic pollutants, most
environmental releases partition to the atmosphere. As a consequence, it is inevitable to adapt effective technologies for
the treatment of industrial VOC emissions, along with the more strict emission regulations in China.

Waste gas treatment processes using bioreactors have gained popularity in control of VOCs and odour, since they offer a
cost effective and environment friendly alternative to conventional air pollution control technologies. However,
industrial VOC emissions usually contained several recalcitrant pollutants (e.g., xylenes, toluene and mercaptans
realased from the pharmaceutical industry), making it difficult for a conventional bioreactor to efficiently remove all of
the target pollutants and strictly meet discharge limits in force. Therefore, one additional pretreatment step is required
for removing these recalcitrant pollutants emitted from the industrial processes. Due to the high oxidation potential and
reactivity of ozone and other reactive species, non-thermal plasmas technology has been widely used for removing
recalcitrant pollutants from industrial VOC emissions. Nowadays, combination of a bioreactor with some physicalchemical
approaches has attracted increasing attention in order to overcome the weaknesses of biodegradation-alone
systems.

In this study, an innovative design of biotreatment system for the removal of the recalcitrant compounds by integrating
pretreatment steps of non-thermal plasmas oxidation and water absorption with a biotrickling filter was studied. In this
integrated process, a dielectric barrier discharge (DBD) reactor was adopted as a pretreatment step in order to transfer
these recalcitrant compounds to easily biodegradable compounds. Furthermore, water absorption followed with the DBD
reactor was designed to keep the stability of a biotrickling filter from the side-effect of overdose ozone, and
simultaneously enhance the absorption of target pollutants into liquid medium. As the key process for the removal of
VOCs and odorous compounds, a biotrickling filter inoculated with two new isolates (Pandoraea sp. WL1 and
Pseudomonas sp. WL2) for removing a mixture of two model pollutants (p-xylene and ethyl mercaptan), was acclimated
to steady-state operation. In this process, the integrated BTF system with a dielectric barrier discharge (DBD) reactor as
the main pretreatment step showed a better performance (removal efficiency of 70%~100%) than that of the BTF-alone
system (removal efficiency of 58%~77%) with total VOC concentrations of 300~1500 mg/m3 (1:1 ratio for ethyl
mercaptan paired with p-xylene). Key interactions identified include inhibition of p-xylene degradation and
enhancement of ethyl mercaptan degradation for a paired mixture relative to single substrate degradation. These indicate
that this novel bioreactor could effectively overcome these operational limitations for poorly soluble and recalcitrant
pollutants in the industrial application.

Abstract

Microwave (MW) uses electrical energy. A catalytic material added MW hydrolysis is an effective
way of reducing electricity consumption and enhancing the hydrolysis of cattle manure. In this study,
catalytic materials, such as H2SO4 as the strong acid and NaCl as the ionic material, were applied to
enhance the hydrolysis of cattle manure. Food wastewater (FWW) was also applied as the ionic
material because of its high contents of NaCl and other ions such as K+, NH4+, NO3-. After catalysts added MW-assisted
thermo-chemical hydrolysis, the maximum increases of SCODrelease/TCODinitial ratio were 0.064 for H2SO4, 0.050 for
NaCl and 0.068 for FWW. And the SCODrelease/TCODinitial ratio of FWW addition was 1.4 times higher than that with
NaCl addition at a MW power of 800 W and target temperature of 40°C. The specific SCOD increase per energy
consumption (mgSCOD/kJ) was 169.6 for H2SO4, 132.5 for NaCl and 180.2 for FWW. In biochemical methane
potential tests, methane productions of H2SO4 and FWW addition were 1.25 times and 1.30 times higher than that with
MW only assisted hydrolysis. Therefore, FWW was found to be good agent for enhancing the MW efficiency.

Department of Civil and Environmental Engineering, Pusan National University, South Korea

Abstract

Microwave (MW) is a promising method for the solubilization of waste activated sludge (WAS) due
to the accelerated reaction rates, environmental friendliness and low overall cost compared to
conventional heating. In this study, several chemical agents, including H2SO4, NaCl and food
wastewater, were applied to enhance the MW effect on the solubilization of WAS. NaCl had a
positive effect on WAS solubilization compared to that of the MW only-assisted pretreatment by 30.1%. Therefore, food
wastewater was used to enhance the MW efficiency in WAS solubilization owing to its high NaCl content. Food
wastewater also improved the WAS solubilization efficiency by 60.3%. In the experiment of H2SO4-assisted MW
pretreatment, the WAS solubilization efficiency was improved by 50.6% compared to that by MW-only assisted
pretreatment. After the WAS pretreatment by microwaves with the chemicals, a biochemical methane production (BMP)
test was conducted. In these experiments, the amount of biogas produced from WAS pretreated with NaCl, food
wastewater and H2SO4 assisted microwaves was 11.3, 31.6 and 29.7% higher, respectively, than that of the nonpretreated
WAS. Therefore, the most appropriate chemical is food wastewater because of solubilization efficiency as
well as waste reuse, cost and environmental friendliness.

Department of Civil and Environmental Engineering, Pusan National University, South Korea

Abstract

Microwave (MW) is a promising method for the solubilization of waste activated sludge (WAS) due
to the accelerated reaction rates, environmental friendliness and low overall cost compared to
conventional heating. In this study, several chemical agents, including H2SO4, NaCl and food
wastewater, were applied to enhance the MW effect on the solubilization of WAS. NaCl had a
positive effect on WAS solubilization compared to that of the MW only-assisted pretreatment by 30.1%. Therefore, food
wastewater was used to enhance the MW efficiency in WAS solubilization owing to its high NaCl content. Food
wastewater also improved the WAS solubilization efficiency by 60.3%. In the experiment of H2SO4-assisted MW
pretreatment, the WAS solubilization efficiency was improved by 50.6% compared to that by MW-only assisted
pretreatment. After the WAS pretreatment by microwaves with the chemicals, a biochemical methane production (BMP)
test was conducted. In these experiments, the amount of biogas produced from WAS pretreated with NaCl, food
wastewater and H2SO4 assisted microwaves was 11.3, 31.6 and 29.7% higher, respectively, than that of the nonpretreated
WAS. Therefore, the most appropriate chemical is food wastewater because of solubilization efficiency as
well as waste reuse, cost and environmental friendliness.

PURIFICATION AND CHARACTERIZATION OF XYLANASE FROM BACILLUS PUMILUS STRAIN K-22 ISOLATED FROM WATER SAMPLES

Abstract

A bacterium, designated as strain K-22 was isolated from GaramChashma, Manghopir, and Karachi that produced xylanase enzyme when cultured on nutrient agar supplemented with birch wood xylan, as indicated by clear zone of hydrolysis (2.0 cm) around its growth. The bacterium was identified as Bacillus pumilus with 99% similarity with type strain D4H3. Strain K-22 produced maximum amount of enzyme at 37°C, pH 8, 0.4% xylan concentration, 3% inoculum and 2% salt concentration. Xylanase enzyme was purified to homogeneity by column chromatography as indicated by a single protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with size of approximately 78 kDa. The enzyme was stable at 20 to 70°C (with an optimal temperature 50°C) and in a pH range of 5.0-10 (with an optimal pH of 8.0). It was stable in the presence of various metal ions such as Mn+2, Ca+2, and Mg+2 while Zn+2, Cu+2 and Hg+2 inhibited the activity. Xylanase activity was also inhibited by chemical reagents such as EDTA, SDS, PMSF, and Marcaptoethanol while NaCl and Sodium azide had very little effect on xylanase activity. Hence it is concluded, that xylanase purified from Bacillus pumilus strain K-22 can be useful in various industrial applications.

HIGHLY EFFICIENT GENOME EDITING FOR CROP IMPROVEMENT

Abstract

The burgeoning demand for plant-derived products, such as food, feed, fuel and fiber, underlies the
importance of methods to continuously improve crop varieties with higher yields, lower input costs
and better nutrition value. Recent advances in precise genome editing open up new opportunities to
develop novel crop varieties with valuable traits. Precise genome editing often require targeted
cleavage of specific chromosomal sequences, which generates single or double strand DNA breaks
and activates endogenous DNA repair pathways. To date, four classes of programmable sequence-specific nucleases,
meganuclease (homing endonuclease), zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases
(TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPRS-associated protein 9
(Cas9) nuclease, have been developed to cleave almost any sequences in any species. In this study, TALEN technology
is employed to edit complex crop genomes. High frequency of gene editing events have been identified from a number
of crop species, such as soybean, potato and canola. High value traits generated through this technology and their
regulatory status will be presented.

Abstract

Activated carbon derived from sargassum(SAC), normally disposed as waste, has been examined as
potential catalysts for selective catalytic reduction (SCR) of NO with NH3 in the temperature range
of 50-250 °C. The influence of preparation methods, phosphoric acid impregnation ratios and surface
nitrogen functional groups, were investigated. An array of analytical techniques, including BET,
SEM-EDX, EA, FTIR and XPS, were also employed to study the structural properties, elemental content, and
distribution of nitrogen-containing groups of the catalyst prepared.

The N-modified SAC samples presented higher catalytic activity than the virgin SAC samples. The N-doped commercial
activated carbon(NCAC) achieved a maximum NOx conversion of 75% at 150 °C and then decreased to 55% at 250 °C.
NOx conversion was maintained above 75% and achieved a maximum of 87% at 150 ⁰C for the NSAC. Besides, N2
selectivity was maintained above 95%. In conclusion, the activated carbons prepared from sargassum could be used as
an alternative of the commercial AC. Moreover, the modification with urea on the SAC leads to the increase of its SCR
activity and N2 selectivity.

CONFERENCE ABSTRACT (IN PROCESS)

Plant and Environment (Track)

College of Chemical and Biological Engineering, Zhejiang University, China

Abstract

Biotrickling filter (BTF) applications in two-liquid phase bioreactors (TLPB) are limited by the lack
of agitation, ozone clogging in the long term and a further promotion in mass transfer. Versus a twoliquid
phase BTF with silicone oil as a non-aqueous phase, a two-liquid phase airlift bioreactor
(ALR) was developed to remove gaseous o-xylene. A higher elimination capacity of 26% was
obtained for the two-liquid phase ALR (92 g.m-3.h-1, removal efficiency of 91%) versus the two-liquid phase BTF.
Removal efficiency of the two-liquid phase ALR was constant and above 90% regardless of the inlet concentration.
More complete removal of o-xylene was observed for the two-liquid phase ALR under both steady state and shock-load
conditions. Therefore, sufficient mixing is essential to the TLPBs for the removal of hydrophobic VOCs. However,
microbial diversity in the two-liquid phase ALR was not richer than that of the two-liquid phase BTF. The dominant
microbial community was Comamonas in the two-liquid phase ALR, Chitinophagaceae in the two-liquid phase BTF.

COOPERATION AND COMPETITION OF GLUCOSE AND CATHODE ELECTRONS IN
BIOFILM ELECTRODE REACTOR RELATED TO NOX REMOVAL

Abstract

Nitric oxides (NOx) are hazardous air pollutants that may cause acid rain, acid fog, photochemical
smog, health risks of flora and fauna, etc. More and more attention has been paid to the control of
NOx. Chemical absorption-biological reduction integrated system is one of the promising approaches
which well integrates biological treatment with physicochemical methods.

According to the previous study, cathode electrons could enhance the reduction of Fe(III)EDTA via direct
electrochemical reduction and indirect biological reduction using cathodic hydrogen as electron donor. Moreover, the
denitrifiers could use hydrogen and inorganic carbon source to generate organics which could serve as electron donor
and carbon source.

The reduction features of Fe(III)EDTA using glucose or cathode electrons as sole electron donor were studied
respectively. It is noted that the reaction rate constant with cathode electrons was much higher than that with glucose.
When the concentration of glucose was less than 300 mg/L, the relationship of cathode electrons and glucose was
cooperation, and when the concentration of glucose was more than 300 mg/L, it changed to competition.

CONFERENCE ABSTRACT (IN PROCESS)

Plant and Environment (Track)

THE FIRST EXTENSIVE CASE-STUDY ON THE EFFECTS OF ARTIFICIAL
MYCORRHIZATION OF ACER BUERGERIANUM MIQ. IN HUNGARY

Krisztina Szabo, Vilmos Lovas and Zsolt Erős-Honti

Garden and Open Space Design, Corvinus University of Budapest, Faculty of Landscape
Architecture, Hungary

Abstract

Owing to the increasing environmental disturbances, afforestation of our cities became a difficult
objective. To elaborate liveable environments, the most important mission of experts is to find plant
species and cultivars tolerating urban conditions and to elaborate methods for helping their survival. A possible solution is using artificial mycorrhizae which have manifold beneficial effects for the host
tree. In our research we present the first results on a large-scale investigation on artificially inoculated Acer
buergerianum species in urban domains.

Members of the genus Acer are rarely planted in treelines Acer buergerianum is one of the most popular trees in Easter
Asian cities and it was included also in the Urban Green Project 2021 as well. Although in Hungary it is found only as
solitaire tree mainly in collection garden, owing to its good tolerance and high decorative value it may have a potential
role in Hungarian urban afforestation.

In our study, endomycorrhizal products available in Hungary were tested on 120 specimens of Acer buergerianum
seedlings. We evaluated the effects of different products and differing inoculation methods. We examined the
colonisation level of the roots and evaluated the vegetative growth intensity of the plants.

Abstract

Transgenic approaches have played significant role for modern plant development. In this regard,
combined gene discovery and functional genomics have proven diversified mechanisms and gene
families, which improved productivity by tolerating different abiotic stresses. We report here genetic
stability and persistent morphological features of transgenic pea (Pisum sativum L.) plants harboring
dicistronic vector construct pG0229MASnhx1/luc in subsequent six generations over a period of five years. In addition
to salt stress tolerance (100mM NaCl), the transgenic plants also showed frost tolerance over wild type (WT) counter
part. The frost tolerance of transgenic pea plants harboring Na+/H+ antiporter from A. thaliana is unexpected yet an
important physiological trait which needs further investigations. The comparison of long term stored transgenic seeds
under 30-50 degree C and glass house grown WT and transformed plants on various morphological and molecular
characterization were investigated. The transgenic plants were found to be morphologically stable and tolerant to NaCl
stress in subsequent generation. Genetic stability of transformed genes was confirmed prior to and after transfer of
transgenic plants in glass house under different climatic conditions. Leaf size, shape and color, plant height, number of
tendrils, flower shape, pod shape and grains were morphologically similar to WT counterpart in all transgenic
generation. The work is in progress.

CONFERENCE ABSTRACT (IN PROCESS)

Marine Biotechnology (Track)

NOVEL CARBON SEQUESTRATION PROCESS BY WHOLE-CELL BIOCATALYSIS:
PARALLEL PRODUCTION OF HYDROGEN AND SYNTHESIS OF LACTIC ACID FROM
DIRECT COUPLING REACTION OF ACETATE AND CO2 BY THE THERMOPHILIC
BACTERIUM THERMOTOGA NEAPOLITANA

Abstract

The heterotrophic bacterium Thermotoga neapolitana produces hydrogen (H2) by fermentation of
organic substrates. The process is referred to as dark fermentation and is completed by synthesis
organic acids whose composition varies in agreement with the culture conditions. Here we show that
dark fermentation is shunted by an unprecedented mechanism of capture and sequestration of carbon
dioxide into lactic acid [1, 2]. This novel pathway is based on direct coupling of CO2 with C2 substrates, such as
exogenous acetate or acetyl-CoA derived from fermentation of carbon-rich materials. In fact, when the bacterial cultures
are supplemented only with carbohydrates, synthesis of lactic acid occurs to the detriment of acetic acid levels but, in
contrast with the currently accepted DF model, synthesis of lactic acid does not affect H2 productivity. On the contrary,
when exogenous acetate is added under CO2-enriched conditions, the bacterial cells perform as microscopic reactors
that are able to synthesize lactic acid through elongation of the acetic acid molecule by addition of carbon dioxide. This
mechanism could represent an unprecedented way of non-photothrophic CO2 fixation in the thermophilic bacterium.
The bacterium can also use bicarbonate but incorporation levels suggest CO2 as substrate for the coupling reaction. The
CO2-dependent fermentation process, which we named capnophilic lactic fermentation, achieves at the same time
production of H2 and conversion of CO2 into added value chemicals without biomass deconstruction [3].

SOME SECONDARY METABOLITES FROM ENDOPHYTES OF THE TURKISH MARINE
SPONGES

Abstract

Endosymbiotic fungus Hypocrea lixii associated with the marine sponge Axinella polypoides,
Penicillium concentricum associated with marine sponge Ircinia oros and Chaetomium globosum associated with Sarcotragus spinulosa were isolated and identified by morfological criteria and
phylogenetic analyses based on internal transcribed spacer (ITS) regions. This paper deals with
isolation of ferulic acid from marine associated fungi Hypocrea lixii, chaetochromin A from marine associated fungi Penicillium concentricum and chaetomin from Chaetomium globosum. Their structures were determined by
comprehensive analyses of 1H and 13C NMRs and LS/MS mass spectrometric datas. Preliminary studies showed that
this is the first report of ferulic acid to be isolated from marine sponge Axinella polypoides associated fungi Hypocrea
lixii. In addition to that, there is no report showed chaetochromin A was isolated from marine fungi Penicillium
concentricum

Abstract

Cysteine (Cys), the first organic sulfur-containing metabolite, serves as a precursor for the synthesis
of glutathione and methionine (Met), two metabolites that are central to plant growth and survival.
Glutathione plays a crucial role in the defence against a wide variety of environmental stresses, while
Met is a protein constituent, and through its first metabolite, S-adenosylMet (SAM), regulates
essential processes required for plant growth. To reveal the relations between glutathione and Met, we used tobacco
plants overexpressing the regulatory enzyme of Met biosynthesis pathway, cystathionine γ-snthase (CGS), and those
overexpressing the yeast gene encoding a feedback-insensitive O-acetylserine (thiol)lyase (OASTL) in the plastids and
in the cytosol that regulate the levels of Cys and glutathione. We crossed between the two transgenic lines to determine
that the level of Met can significantly increase in plants overexpressing the plastidic OASTL with AtCGS, accompanied
by a reduction in glutathione. The results strongly suggest that the flux towards Met is relatively high, and thus Met can
be considered as an intermediate metabolite in the pathways leading to its various associated metabolites. In addition,
flux and metabolic profiling analyses indicated the existence of metabolic competition between the biosynthesis
pathways of Met and glutathione on their common precursor, Cys, and that this competition is more crucial under
oxidative conditions when more Cys is required for the synthesis of glutathione. Plants overexpressing AtCGS with or
without the yeast enzyme were significantly more sensitive to oxidative stress, indicating the reason why the levels of
Met remained low during the evolution.

Pharmaceutical Biotechnology

College of Natural and Applied Sciences, Crescent University, Abeokuta, Nigeria

Abstract

Hepatitis is one of the major health problems in Nigeria which sometimes may lead to even death.
Nigeria has the treasure of medicinal plants, which may be the best source of remedies for the
treatment of liver diseases. Thus identification of a potential therapeutic agent for the protection of
liver from the hepatotoxins will provide a useful way for the prevention of liver related sickness. Our
studies identified a plant with full potential of hepatoprotective activity. The etanolic extract of Annona muricata leaves
were screened for its hepatoprotective activity in paracetamol induced liver damage in Albino rat at a dose of 200 mg/kg
bw. The ethanolic extract exhibited a significant protective effect by lowering serum levels of glutamic oxaloacetic
transaminase, glutamic pyruvic transaminase, alkaline phosphatase, total bilirubin, increasing the antioxidant enzymes
(superoxide dismutase, catalase, peroxidase, glutathione reductase and glutathione-S-transferase as well as glutathione
level. Liv.52 was used as positive control. The effect of the drug was observed by changes in serum marker ALT, AST,
ALP, total serum protein and bilirubin levels. The extract did not show any mortality up to a dose of 1000g/kg bw.

Abstract

The recent discovery of the phenomenon of RNA interference has led to it’s application in various
aspects of plant improvement. Crops can be modified by engineering novel RNA interference
pathways that create small RNA molecules to alter gene expression in crops or plant pests. RNA
interference can generate new crop quality traits or provide protection against insects, nematodes and
pathogens without introducing new proteins into food and feed products, this is an advantage in contrast with
conventional procedures of gene transfer. RNA interference has been used to develop crop varieties resistant to diseases,
pathogens and insects. Male sterility has been engineered in plants using RNA interference. Better quality crops have
been developed through the application of RNA interference etc. The objective of this paper is to highlight the
application of RNA interference in crop improvement and to project it’s potential future use to solve problems of
agricultural production in relation to plant breeding.

EFFICACY OF SOME MANGROVE METABOLIC AGENTS AGAINST CYCLOXYGENASE, 5-
LIPOXYGENASE AND ACETYLCHOLINESTERASE ENZYMES

Abstract

Mangroves are highly productive ecosystem with various important economic and environmental
functions. Mangroves are the key elements in marine environment, they diverse metabolites to adapt
with the requirement of their challenging ecosystem. This make them an interesting source for natural
bioactive metabolites.

Inflammation is a complex disorder which involve biosynthesis of certain type of prostaglandins that are responsible for
the sensation of pain.

The primary enzyme responsible for prostaglandin synthesis is cylooxygenase (COX) which occurs in two isoform
cyclooxygenase-1, 2. Inhibition of prostaglandin produced by cyclooxygenase-2 (COX-2) is a target. However, some of
the developed selective COX-2 drugs showed serious side effects on clinial trials.

Recent evidence support the search for antiinflammatory agents from natural origin based on dual inhibitors of COX and
5-lipoxygenase (LOX), an enzyme in leukotriene biosynthesis. leukotrienes was suspected to play a role in
Gastrointestinal toxicity observed with some of the previously approved drugs.

In this study, we investigated COX and LOX, and AchE inhibitory effects of some Malaysian mangrove plants based on
their ethnobotanical uses for the traditional treatment of inflammatory related ailments. Plant metabolites appeared to
possess dual activities are currently unders investigation in our laboratory for their possible role in suppression of
fibroblast-like synoviocytes in rhumatoid arthritis.

CONFERENCE ABSTRACT (IN PROCESS)

Pharmaceutical Biotechnology

Abstract

The human cannabinoid receptor CB2 belongs to the class A of heptahelical G protein-coupled
receptors (GPCR) and is an attractive target for the development of drugs for management of pain,
inflammation, osteoporosis and treatment of immunological disorders. High resolution structural
studies are critical to obtain insights into the molecular mechanisms of ligand binding and activation
of CB2.

We developed methods for expression in milligram quantities, purification, reconstitution in lipid bilayers and
stabilization of the functional recombinant CB2 as well as efficient stable isotope labeling of CB2 by high density
fermentation were developed, enabling NMR studies. NMR analysis of labeled receptor reconstituted in proteoliposomes
in agonist or inverse-agonist-bound form will be reported.

Monoclonal antibodies were raised against the purified CB2, and the affinity of interaction with CB2 was
determined by surface plasmon resonance. Finally, we demonstrate the specific effects of lipids with negatively
charged head group in activation of CB2 reconstituted in proteoliposomes as measured by an in vitro G protein
activation which may have important physiological significance as manifested in natural membranes of various
lipid compositions.

CONFERENCE ABSTRACT (IN PROCESS)

Other Areas

ATPASE 6/8 GENE ASSISTED GENETIC VARIABILITY STUDIES AMONG THE POPULATION
OF LABEO ROHITA

Fayyaz Rasool, Shafat Hussain and Shakeela Parveen

Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Lahore-Pakistan

Abstract

Labeo rohita is a commercially important fish of South Asia. A study on the variation amongst the
different populations of on the basis of ATPase 6/8 gene was conducted. The present study evaluates
the potential of complete ATPase 6/8 region of mitochondrial DNA as a marker region to determine
the phylogeography of Labeo rohita from fish farm of region Punjab, Pakistan. mtATPase6/8 (878
bp) regions was used to investigate genetic variation within Labeo rohita and develop a global genealogy of genus Labeo
strains. The mtATPase6/8 region was more variable but the given the wide distribution of Labeo rohita the overall levels
of sequence divergence were low. Levels of haplotype diversity varied widely among countries with Chinese and India
showing the greatest diversity whereas Japanese Labeo had undetectable nucleotide variation. . Chinese and Japanese
carp strains were the most divergent, and their relationships do not support the evolution of independent Asian and
European lineages and current taxonomic treatments. The results revealed that 878 bp of ATPase 6/8 region could be a
promising marker for determining variations at interpopulation as well as intrapopulation levels in Labeo rohita. These
results would facilitate conservation and management of this important species

ARTEMISININ PRODUCTION IN ARTEMISIA ANNUA AND ARTEMISIA DUBIA
FOLLOWING TRANSFORMATION WITH THE ROL ABC GENES AND ELUCIDATION OF
THE SITES OF ITS SYNTHESIS

Abstract

The rol ABC genes have been shown to enhance production of secondary metabolites in plants,
possibly through stimulation of the defense pathway. This report examines the effect of
transformation of A.annua and A.dubia with the rol ABC genes expressing in A. tumefaciens and
A.rhizogenes. The artemisinin content, trichome density and expression of key genes in the
biosynthetic pathway of artemisinin were measured. Artemisinin content was significantly increased in transformed
material of both Artemisia species when compared to un-transformed plants. The artemisinin content within leaves of
transformed lines was increased by a factor of ten, indicating that the plant is capable of synthesizing much higher
amounts than has been achieved so far through traditional breeding. Expression of all artemisinin biosynthesis genes was
significantly increased, although variation between the genes was observed. Cytochrome P450 (CYP71AV1) and
aldehyde dehydrogenase 1 (ALDH1) expression levels were higher than that of amorpha-4, 11 diene synthase (ADS).
Levels of the trichome development and sesquiterpenoid biosynthetic gene (TFAR1) expression were also increased in
all transgenic lines. Trichome density was also significantly increased in the leaves of transformed plants, but no
trichomes were found in control roots or transformed roots. The detection of significantly raised levels of expression of
the genes involved in artemisinin biosynthesis in transformed roots correlated with the production of significant amounts
of artemisinin in these tissues. This suggests that synthesis is occurring in tissues other than the trichomes which
contradict previous theories. This elucidation will help to increase production to meet the increasing demand of
artemisinin because of its pharmacological importance.

Abstract

Total eight endophytic bacteria were isolated from the roots of P. deltoides growing in non
contaminated sites at natural vegetation of Garhwal Himalayas, Uttarakhand, India. Out of these only
SBER3 isolate was able to metabolize wide range polyaromatic hydrocarbons (PAH) and other
hydrocarbon used in the study viz. antracene, napthalene, benzene, chrysene, benzene, toulene and xylene on minimal
salt basal medium (MSB) as sole carbon and energy source. It was identified as Bacillus sp. on the basis of 16s rDNA
sequence. Including this Bacillus sp. SBER3 also produced IAA, phosphate solubilization, siderophore and ACC
deaminase. Along with these traits Bacillus sp SBER3 also inhibited the fungal phytopathogen. Microscopic
examination under the influence of Bacillus sp. SBER3 revealed abnormalities in case of R. solanii, F. oxysporum and
M. phaseolina. However, mean growth rate and survival under varying osmotic stress regime were also evaluated under
vitro condition. Rhizoremediation potential of Bacillus sp. SBER3 was also demonstrated in hydrocarbon amended soil
model system and it showed significant enchancement in shoot length, root length, root and shoot biomass including
stem girth of Populus plant respective to control and concurrently decreased the concentration PAH in soil as estimated
by HPLC analysis. Finally, a multipurpose bioinoculant was formulated by using Bacillus sp. SBER3 isolate in six
different lignocellulosic solid carrier's material for proper bioculture dissemination in bioremediation along with growth
enhancement and protection against deleterious pathogen of P. deltoides in PAH contaminated environment.

CONFERENCE ABSTRACT (IN PROCESS)

Pharmaceutical Biotechnology

WHOLE CELL BIOCATALYST FOR SOYASAPOGENOL B PRODUCTION FROM SOYBEAN
SAPONIN

Hala A. Amin, Mostafa M. Abo Elsoud, Ahmed. F. Sahab

Chemistry of Natural and Microbial products Department, National Research Center, Egypt

Abstract

Soyasapogenol B, aglycone of soybean saponin, is known to have hepatoprotective, antimutagenic,
antivirus, and anti-inflammatory activities. This research examined the use of whole-cell biocatalyst
to produce soyasapogenol B from soybean saponin. It was found that Aspergillus flavus, a fungus
isolated from peanut pods, was capable of expressing extracellular and intracellular saponin
hydrolase enzyme. However, the total enzyme activity produced using fungal whole cells (37U) in the reaction mixture
was about 3times that produced using the extracellular (12.4U) or intracellular (11.5U) enzyme. Cells with maximum
hydrolytic activity for production of soyasapogenol B (12.2 U/g) was obtained using production medium supplemented
by 2% soybean saponin, as inducer for enzyme production, adjusted at pH 9 and incubated at 30°C for 2 days. The
highest yield of soyasapogenol B was achieved when the reaction mixture was incubated at pH 5.5 and 45°C for 48h;
using 20 g wet cells (corresponding to 4% cell dry weight) and soybean saponin (2%, w/v) as a substrate. Under these
optimal conditions, the cells bioconversion efficiency (soysapogenole B yield) increased from 5.3 to 60%. Whole cell
biocatalyst has several advantages with regard to industrial applications: a consistent quality, easy to be prepared and a
very low price compared with purified enzyme. Consequently, this study is significant for production of soyasapogenol
B from soybean saponin on an industrial scale.

ACKNOWLEDGEMENTS
The authors would like to thank for financial support via the tenth research grant (2013-2016) of the National Research
Center of Egypt.

HIGH PRODUCTION OF FATTY ALCOHOLS BY METABOLICALLY ENGINEERED
ESCHERICHIA COLI WITH FATTY ACID STARVATION

Abstract

Microbial synthesis of fatty alcohols from renewable resources has attracted increasing attentions.
However, the low tilters of fatty alcohols hampered its industrialization. Here we designed a novel
strategy for fatty alcohol production based on fatty acid starvation. For the first time, all three acyl-
ACP thioesterases related to formation of fatty acids were knocked out to enhance fatty alcohol
production. The fatty alcohol titer increased about 58%, while fatty acids concentration dropped 73%. Transcriptome
analysis showed that expression levels of genes for fatty acid synthesis and glycolysis pathway were upregulated, while
fatty acid degradation and TCA pathway were downregulated. Furthermore, Fatty alcohol production was enhanced by
deleting the genes responsible for lactate and acetate formation. The optimized strain, E. coli MGL2 with exogenous
fatty acyl-ACP reductase, accumulated fatty alcohols at a remarkable level of 6.33 g/L under fed-batch fermentation.
This is the highest reported titer of fatty alcohols produced by microorganisms.

Abstract

The recognition that different stem cell types can home to tumors following transplantation has
unveiled new possibilities for their use in cancer therapy. Our research is based on simultaneously
targeting cell death and proliferation pathways in tumor cells in an effort to eradicate both primary and metastatic tumors
in the brain using therapeutically engineered stem cells. We have engineered different adult stem cells types to release
therapeutic: (i) pro-apoptotic protein, S-TRAIL (secreted tumor necrosis factor receptor-apoptosis inducing ligand) to
specifically induce apoptosis in tumor cells; (ii) anti-proliferative anti-epidermal growth factor (EGFR) nanobodies
(ENb) to inhibit tumor cell proliferation; (iii) anti-angiogenic thrombospondin (TSP)-1 to target blood vessels supplying
the tumor; and (iv) bimodal anti-proliferative and pro-apoptotic proteins that target both cell proliferation and death
pathways in tumors cells. We have extensively demonstrated the therapeutic efficacy of these engineered stem cells both
in culture and in mouse tumor models. Recently, we have also explored the potential of stem cell loaded oncolytic
viruses for cancer therapy and shown that mesenchymal stem cell (MSC) loaded with different variants of oncolytic
herpes viruses (oHSV) eradicate tumors in mice brains. In an effort to translate these therapeutics into clinical settings,
we have utilized different mouse tumor models generated from our extensively characterized patient derived brain tumor
cells. These mouse models mimic clinical settings of primary tumors and their secondary micro-invasive deposits in the
brain. Inherently linked to the brain tumor therapy paradigm, we have employed imaging markers and optical imaging,
MRI and PET techniques to track stem cells, image apoptosis and changes in tumor volumes in real time in vivo. Within
next 2-5 years, we expect to initiate a clinical study using therapeutic stem cells to treat brain tumor patients and hope to
have a major impact in saving the lives of these patients.

CONFERENCE ABSTRACT (IN PROCESS)

Pharmaceutical Biotechnology

DEVELOPMENT OF NEW ANTIMICROBIAL AGENTS OF INORGANIC ORIGIN

Abstract

In the recent years, due to the development of new strains, resistance of bacteria to the commonly
used antibiotics drug has increased. Most of the Antibiotics being used, today, are of organic nature.
Some of these are extremely irritant and even toxic, therefore, there is much interest in finding ways
to formulate new types of safe and cost-effective biocidal materials. Nanosize inorganic material
such as metals and their salts during their antibiotic action, can cause denaturation of proteins present in bacterial cell
walls. This paper will highlight the synthesis, characterization and antimicrobial activity of as-synthesized inorganic
antibiotics against gram positive (Staphylococcus aureus and Streptococcus) as well as gram negative (Pseudomonas
aeruginosa and Escherichia coli) bacteria using paper disc diffusion technique. Effects of doping silver and Sulphur in
ZnS nanoparticles on their antimicrobial activity will be reported and discussed. The antimicrobial performance of assynthesized
nanomaterial will be compared with the commonly used antibiotic drugs.

TERMITE CONTROL BIOTECHNOLOGY

Abstract

Termites are considered as important pests that could cause severe wood damage and economic loses
in urban, agriculture and forest of Malaysia. The ability of termites to degrade cellulose depends on
association of gut cellulolytic microflora or better known as mutual symbionts. With the idea of
disrupting the mutual symbiotic association, better pest control practices can be attained. This study is
aimed to isolate cellulolytic bacteria from the gut of termites and carry out antibacterial studies for termite. Confirmation
of cellulase activity is done by qualitative and quantitative methods. Impacts of antibiotics and their combinations as
well as heavy metals and disinfectants are conducted by using disc diffusion method. Effective antibacterial agents are
then subjected for termite treatment to study the effectiveness of the agents as termiticides. 24 cellulolytic bacteria are
isolated, purified and screened from the gut of termites. All isolates were identified as Gram-negative with either rod or
cocci in shape. For antibacterial studies result, isolates were found to be 100% sensitive to 4 antibiotics (rifampicin,
tetracycline, gentamycin and neomycin), 2 heavy metals (cadmium and mercury) and 3 disinfectants (lactic acid,
formalin and hydrogen peroxide). 22 out of 36 antibiotic combinations showed synergistic effect, while 15 antibiotic
combinations showed antagonistic effect on isolates. The 2 heavy metals and 3 disinfectants that showed 100%
effectiveness as well as 22 antibiotic combinations that showed synergistic effect were used for termite control. Among
the 27 selected antibacterial agents, 12 of them were found to be effective to kill all the termites within 1 to 6 days.
Mercury, lactic acid, formalin and hydrogen peroxide were found to be the most effective termiticides in which all
termites were killed within 1 day only. These effective antibacterial agents possess a great potential to be a new
application to control the termite pest species in the future.

CONFERENCE ABSTRACT (IN PROCESS)

Medical Biotechnology (Track)

NEW BIO-TECHNOLOGY TOOLS MAY DEFEATS DIABETES MELLITUS

Sultan Ayoub Meo

Physiology, College of Medicine, King Saud University, Riyadh, KSA

Abstract

Diabetes mellitus is a life-long disease and is swiftly increasing in all age groups and both genders. It
involves various physiological functions, organs and multiple systems, and is associated with wide
ranging and devastating health complications. New figures have broken all the previous prevalence
records, global prevalence of diabetes is 8.3% which means that 382 million adults are diabetic, and
the number is expected to rise to 592 million by 2035. Currently, six countries in the Middle East including Saudi
Arabia, Bahrain, United Arab Emirates, Kuwait, Oman and Egypt are among the world's highest for the prevalence of
diabetes. Management of Diabetes Mellitus presents challenges to the health care providers. In the 21st century, there
has been constant progress in the management of diabetes mellitus. The introduction of new insulin delivery systems and
glucose monitoring devices has enhanced the ability of both patients and medical teams to better define and develop the
therapeutic strategies. Recent advances in devices for insulin administration and glucose monitoring having a profound
effect on the lives of diabetic patients. It is strongly suggested that, the science community, physicians must adopt the
new bio-technology tools in the management of diabetes mellitus to defeat this debilitating disease.

Abstract

Phytophthora species belong to a unique class of eukaryotic microorganisms called oomycetes that
are morphologically similar to but phylogenetically distant from true fungi. Phytophthora species
cause enormous crop losses and damages to natural ecosystems. For example, the Irish Famine causal
agent Phytophthora infestans has still been a worldwide major threat to sustainable potato production,
owing largely to capability of the pathogen in erasing efficiency of disease resistance genes. Disease resistance is usually
achieved by plant recognition of pathogen effectors. Disruption of such recognition, more frequently by genetic or
epigenetic changes of expression of recognized effector genes in the pathogen, leads to loss of disease resistance. In this
report, we describe an approach for engineering disease resistance in plants against Phytophthora pathogens. Plants
encode small secreted proteins in response to stresses, which potentially play a function in triggering resistance, similar
to pathogen effectors. Global striking changes in host plant gene expression are well documented in response to
pathogen infection. We show that engineered resistance genes based on combinations of a plant cis element highly and
specifically responsive to Phytophthora infection and small defense related protein genes lead to high level of resistance
to infections by Phytophthora pathogens. The engineered disease resistance to broad-spectrum and is potentially
durable. Our results show a novel approach for exploring plant endogenous small proteins in engineering durable and
broad-spectrum plant disease resistance against pathogens.

CONFERENCE ABSTRACT (IN PROCESS)

Plant and Environment (Track)

IDENTIFICATION OF A MAJOR LOCUS CONTROLLING NA+ UPTAKE IN RICE (ORYZA
SATIVA)

Muhammad Saeed, Md. Shah Kamal and Javed Iqbal

Department of Botany, Government College University, Faisalabad, Pakistan

Abstract

Climate change is adversely affecting World crop production. Climate change has intensified
already existing menace of soil salinity in most of the countries. Na+ uptake is an important
attribute with respect to salinity tolerance in plants. Plants which manage to keep low
concentrations of Na+ in their internal environment, relative to the external environment
surrounding their roots, perform better under saline conditions. Quantitative trait loci (QTL)
involved in salinity tolerance in rice (Oryza sativa) were identified by using an F2 population derived from the
cross of IR - 36 (salt sensitive) and Pokkali (salt tolerant). Plant material was evaluated under normal and saline
conditions and data for biochemical traits and grain yield were collected at the maturity stage. A major locus
controlling Na+ uptake was identified on chromosome 7 with a phenotypic variance explained (R2) value of
72.57%. This was a novel QTL not reported previously. Associated marker with this QTL was RM248. Allele
from salt tolerant parent, Pokkali, was responsible for an increase in Na+ uptake. This QTL may be a good
candidate for marker-assisted selection (MAS) to develop salt tolerant rice cultivars with potential to give stable
yield under climate change scenario.

Abstract

In Morocco, fisheries are very diversified. However, the sources used are dominated by small
pelagics. According to FAO, Morocco's fisheries production in 2014 amounted to one million tonnes.
This record is due to a single species, sardine (Sardina pilchardus), which Morocco is both the largest
producer and exporter worldwide.

The establishment of a regulatory environmental policy increasingly strict encourages Moroccan manufacturers to
consider waste / by-products resulting from the processing of fish. The search for biologically active molecule extracted
from these co-products begins to generate interest. It seems interesting, as part of the new strategy of development of the
fisheries sector in Morocco called "Halieutis Plan" and the development of a National Charter for Environment and
Sustainable Development, to develop a new technique to guide enhancement of marine by-products into high value
products rather than considering them as waste low interest.

It is in this context that the present project is part of the value scales of sardine (Sardina pilchardus) to reduce the
pollution load discharged and recover valuable biomolecules with high added value, the collagen in this case.

The preliminary results of this study demonstrated the ability to produce, in the pre industrial scale, collagen powder
form sardine scales with a yield of 26%. This collagen contains a large amount of 78% protein and a medium amount of
ashes 16.8%. Regarding the amount of water, it represents only 9.5%. Biomass scales representing 2% of the weight of
the fish and is not even valued, can become raw material for the production of collagen used for health nutrition,
pharmaceutical, cosmetic and biomedical.

CONFERENCE ABSTRACT (IN PROCESS)

A STUDY ON AGING KINETICS, INHIBITION AND REACTIVATION OF ORGANOPHOSPHATE COMPOUNDS WITH QUAIL'S AND CHICK'S CARBOXYLESTERASE

Kasim Sakran Abass
School of Public Health, Faculty of Veterinary Medicine, University of Kirkuk, Kirkuk, Iraq

Abstract

Organophosphate inhibits carboxylesterase thus regular treatment includes carboxylesterase oxime reactivator in combination with anti-muscarinic agents. In order to progress the understanding of species differences and to enable an additional reliable extrapolation of animal result to quail a study was initiated to inspect the effect of insecticide compounds, i.e. mevinphos, ethoprop and fenthion, with quail’s and chick’s serum carboxylesterase. The rate constants for the inhibition of carboxylesterase by these organophosphate and for the aging and spontaneous reactivation of organophosphate-inhibited carboxylesterase enzyme as well as for the oxime-induced reactivation of organophosphate-inhibited carboxylesterase enzyme by the oximes trimedoxime, methoxime, N,N'-(ethano)bis(4-hydroxyiminomethyl)pyridinium methanosulphonate, HI-6 and HLö-7 were determined in this study. Compared to serum quail carboxylesterase and chicks carboxylesterase displayed a lower sensitivity towards the inspected organophosphate. Furthermore, a slower aging and spontaneous reactivation of serum quail carboxylesterase enzyme was recorded. The potency of the detected oxime reactivator was remarkably lower with organophosphate -inhibited serum quail carboxylesterase.

Abstract

The high-molecular fractions from the species of two genera (Symphytum and Anchusa) of Boraginaceae family Symphytum asperum, S. caucasicum, S. officinale and Anchusa italica were isolated. According to 13C, 1H NMR and 2D heteronuclear 1H/13C HSQC spectral data the main structural element of these preparations was found to be a regularly substituted polyoxyethylene, namely poly[3-(3,4-dihydroxyphenyl)glyceric acid] (PDPGA) or poly[oxy-1-carboxy-2-(3,4-dihydroxyphenyl)ethylene]. PDPGA represents a new class of natural polyethers with a residue of 3-(3,4-dihydroxyphenyl)glyceric acid as the repeating unit. Most of the carboxylic groups of PDPGA from A. italica unlike the polymer of Symphytum species are methylated. Such caffeic acid-derived biopolymer to our knowledge has not been known and has been identified for the first time. According to results of in vitro and in vivo experiments PDPGA could be considered as potential antiinflammatory, wound healing and anti-cancer therapeutic agent.

PDPGA exerted anti-cancer efficacy in vitro and in vivo against androgen-dependent and -independent PCA cells via targeting androgen receptor, cell cycle arrest and apoptosis without any toxicity, together with a strong decrease in PSA level in plasma.

Overall, this study identifies PDPGA as a potent agent against PCA without any toxicity, and supports its clinical application.

CONFERENCE ABSTRACT (IN PROCESS)

USING CITRULLUS COLOCYNTHIS CORTEX AS NEW MEDICINAL HERB TO TREAT GOUT

Abstract

The present study aims to investigate the methanolic extracts effect of Citrullus colocynthis cortex on xanthine oxidase. It is also known for its highly bitter taste such as 'bitter cucumber'. Cortex of such herb tested in vitro at concentration (100 µg/ml) for their inhibition potencies expressed as % inhibition of XO activity. The test plant inhibition activity of xanthine oxidase found to be (78.30±0.92 %) compared with Allopurinol (positive inhibitor) (p < 0.05). The in vivo hypouricaemic activity of such extract (dose of 0.5 mg/kg b.w.) using oxonate-induced hyperuricaemic mice were tested for their ability to reduce the serum urate level and inhibitory actions on the XO enzyme activities in the mouse liver. Potassium oxonate (150 mg/kg, i.p.), an uricase inhibitor was used to induce hyperuricaemia. Allopurinol (10 mg/kg, p.o.) was used as the positive control. The fractions when administered to hyperuricaemic mice produced a significant reduction in serum urate levels due to inhibitory actions on the XO enzyme activities in the mouse liver. Phytochemical screening of the Citrullus colocynthis cortex revealed the presence of tannins, flavonoids, alkaloids and terpenoids which may be partly responsible for the beneficial effect on hyperuricaemia and gout.These results suggest that fractions of Citrullus colocynthis cortex could be used as a potential source to treat gout and other inflammatory disorders.

CONFERENCE ABSTRACT (IN PROCESS)

1Department of Pharmacology, College of Medicine and KHUH, King Saud University, Kingdom of Saudi Arabia
2Department of Anatomy & Histology, College of Medicine and KHUH, King Saud University

Abstract

Lung fibrosis is a common side effect of the chemotherapeutic agent, bleomycin. Current evidence suggests that reactive oxygen species may play a key role in the development of lung fibrosis. The present work studied the effect of green tea extract on bleomycin–induced lung fibrosis in rats. Animals were divided into three groups: (1) Saline control group; (2) bleomycin group in which rats were injected with bleomycin (15mg/kg,i.p.) three times a week for four weeks; (3) bleomycin and green tea group in which green tea extract was given to rats (100mg/kg/day, p.o) a week prior to bleomycin and daily during bleomycin injections for 4 weeks until the end of the experiment. Bleomycin–induced pulmonary injury and lung fibrosis that was indicated by increased lung hydroxyproline content, elevated nitric oxide synthase, myeoloperoxidase (MPO), platelet activating factor (PAF), tumor necrosis factor α (TNF_α ), transforming growth factor 1ß ( TGF1ß ) and angiotensin converting enzyme (ACE) activity in lung tissues. On the other hand, bleomycin induced a reduction in reduced glutathione concentration (GSH). Moreover, bleomycin resulted in severe histological changes in lung tissues revealed as lymphocytes and neutrophils infiltration, increased collagen deposition and fibrosis. Co-administration of bleomycin and green tea extract reduced bleomycin–induced lung injury as evaluated by the significant reduction in hydroxyproline content, nitric oxide synthase activity, levels of MPO, PAF & TNF-α & ACE in lung tissues. Furthermore, green tea extract ameliorated bleomycin– induced reduction in GSH concentration. Finally, histological evidences supported the ability of green tea extract to attenuate bleomycin–induced lung fibrosis and consolidation. Thus, the finding of the present study provides that green tea may serve as a novel target for potential therapeutic treatment of lung fibrosis.

Abstract

The present study investigated the analgesic and anti-inflammatory effects of chlorogenic acid (CGA), a polyphenolic compound present in many foods and beverages using carrageenan (Carr)-induced paw edema in rats and formalin-induced algesia in mice. The molecular mechanisms of CGA were studied through induction of endogenous & exogenous superoxide anion stress in the aforementioned rodent inflammation & algesia models. The results revealed that treatment of rats with CGA (50, 100 or 150 mg/kg) significantly reduced the rats paw edema induced by carrageenan and the formalin- induced pain in mice (P<.05) as compared to control groups. A significant reduction in rat paw volume in nitric oxide induced edema was observed (P<.05), also CGA produced a significant reduction in malondialdehyde (MDA) and a significant increase in reduced glutathione (GSH) in paw tissues (P<.05). These results confirm that CGA has both analgesic and anti-inflammatory properties which may be related to the ability of this polyphenol to reduce the levels of superoxide and peroxynitrite anion radicals. The results of the present work showed that CGA represents a promising potential drug of natural anti-inflammatory property for the development of new drugs that may help to control oxidative stress and consequently the inflammatory response.

APPLICATION OF NEWER MOLECULAR METHODS FOR DIAGNOSING AND DRUG RESISTANCE DETECTION IN MYCOBACTERIUM TUBERCULOSIS: EVALUATION IN INDIAN SETTING

Abstract

Tuberculosis is global health problem, specially after AIDS epidemic. India is having largest pool of drug resistant tuberculosis. The most important concern is its early diagnosis and drug resistance. The conventional methods of diagnosis and drug resistance detection are cumbersome, takes several weeks and lack reproducibility. Therefore in recent years several molecular methods such as conventional monoplex and multiplex polymerase chain reactions, real time PCR assays, Line probe assay and Xpert MTB/RIF have been made commercially available. The MTBDRplus line probe assay (LPA) and Xpert MTB/RIF have been endorsed by World Health Organization and both these tests can be used for detecting Tuberculosis as well as for drug resistant detection in Mycobacterium tuberculosis. However, there is no clarity regarding the superiority of one over the other. Therefore, for the first time from India, we carried out a prospective study, to evaluated the efficacy of Xpert MTB/RIF and LPA on culture confirmed samples. A total of 405 sputa of suspected drug resistant tuberculosis patients were included. Of these, 285 samples were smear positive and all these were subjected to LPA. Seventy-two (25.8%) samples showed multi-drug resistance, 62 (22.2%) showed rifampicin monoresistance, 29 (10.3%) isoniazid monoresistance and 116 (41.5%) were pan-susceptible. All 62 rifampicin monoresistant samples detected by LPA were tested by Xpert MTB/RIF using cartridge version G4. Of these, 38 (61.4%) showed concordance with LPA showing rifampicin resistance while 21 (33.8%) were found discordant susceptible to rifampicin by Xpert MTB/RIF using cartridge version G4. Of the 116 pan-susceptible samples, only 83 were available for Xpert MTB/RIF testing; of which 4 (5.1%) were found rifampicin resistant, 74 (94.8%) were susceptible. The 25 discrepant samples were further subjected to MGIT960 drug-susceptibility testing. The MGIT960 results showed 100% agreement with LPA results but only 64.4% agreement with Xpert MTB/RIF results. Sequencing analysis of discrepant samples showed 91.3% concordance with LPA but only 8.7% concordance with Xpert MTB/RIF assay. These isolates were characterized by spoligotyping and mycobacterial interspersed repetitive-unit–variable-number-tandem-repeat (MIRU-VNTR) analysis. Interestingly, most of these strains were CAS (CAS-1_Delhi [SIT 26] and 2 of SIT 846), followed by Orphan (SIT 27), and only 1 belonged to the MANU2 (SIT 1976) genotype. NO isolate was Beijing type.

This study shows that Xpert MTB/RIF may not be the first choice of molecular test to be used as point of care as primary screening test.

MICROSPORIDIA AND ZINGIBER OFFICINALE (GINGER); AN EMERGING PARASITE AND
AN OLD MEDICINAL PLANT: IN VIVO TRIAL.

Abstract

Microsporidia, of the genus Enterocytozoon, are an important cause of life-threatening diarrhea
especially in immunocompromised hosts. There are controversies on the use of albendazole in
treatment, whereas, fumagillin was to be more effective but with undesirable side effects. Ginger has
been used as an antimicrobial agent since ancient times. However its potential therapeutic effect
against Enterocytozoon bieneusi has not been tested. This study was done to investigate the effect of ginger as a
prospective therapy for microsporidia versus fumagillin in immunocompetent and immunosuppressed mice. Also, to
report the synergistic effect of the two compounds together in a drug-combination regimen. Enterocytozoon bieneusi
was the species identified in the stool samples collected from immunocompromised patients and was used to initiate the
in vivo infection in albino mice. Animals were divided into three major groups. Group I: Normal, non-infected nontreated,
control group; group II: infected, immunocompetent group; and group III: infected, immunosuppressed group.
Each infected group was subdivided into four equal subgroups a, b, c and d which comprise non-treated, fumagillintreated,
ginger-treated, and combined ginger/fumagillin treated mice respectively. Evaluation of the ginger efficacy in
infected mice was achieved by assessment of fecal spore shedding, intestinal spore load, and biochemical assay which
aimed at estimation of the malondialdehyde level and total antioxidant capacity. Spore count in both stool and intestinal
sections and malondialdehyde level decreased significantly with ginger treatment. Best results were obtained when
ginger is combined with fumagillin in all measured parameters. Ginger could be a good enhancer for fumagillin efficacy
to eradicate infection. However, further studies on their its principles, mechanisms of action, toxicity evaluation are still
needed.

Abstract

Aim: Hepcidin is a key regulator of the iron homeostasis. The pathogenesis of the anemia of chronic
disease (ACD) is coupled with hepcidin stimulated synthesis as a result of inflammation. Hereditary
hemochromatosis covers genetic disorders in the metabolism of iron by the appearance of superimposition.
Iron overload leads to organ damage such as liver fibrosis, cirrhosis, cardiomyopathia. In
patients with iron-deficiency anemia (IDA) and those with low serum ferritin latent and pre-latent form of iron
deficiency without concurrent anemia (due to blood loss or insufficient iron intake from food) sets very low hepcidin
levels. The purpose of our study was to investigate serum hepcidin in patients with ACD and IDA in different diseases in
order to help finding the right therapeutic choice.

Discussion: Reliable method for serum hepcidin is important for the proper therapeutic approach in patients with ACD
and IDA in different diseases. The rate of rise may differentiate patients with functional iron deficiency than those with
reticuloendothelial blockade. Monitoring of serum hepcidin concentrations in parenteral therapy with iron preparations
reduces the risk of toxic overload of iron in the body. Hepcidin is a potential indicator of iron deficiency in RA patients
with anemia and an active inflammatory process.

CONFERENCE ABSTRACT (IN PROCESS)

Biological Sciences, Indian Institute of Science Education and Research Kolkata, India

Abstract

Plants face a plethora of biotic stresses in their agro-ecological environments. Responses of plants
tailored to these stresses involve perception, processing and integration of external information into
cellular and physiological machinery. This involves elicitation of complex signaling networks. But
hese signaling networks remain poorly characterized in crop species such as wheat. Moreover, other
than the involvement of some transcription factors (TFs), how cellular signaling is modulated during attack of fungal
pathogens and herbivores remains poorly understood even in model plants. On the other hand, small regulatory RNAs
(smRNAs), such as microRNAs, have appeared as master regulators of cellular signaling events in processes such as
development and differentiation. Our functional studies on components small RNA machinery suggest that smRNA
biogenesis pathways have evolved in specialized manner during adaptation to specific stresses. Use the Argonaute
(AGO) proteins (the central component of the smRNA pathways) as candidates for studying evolution of smRNA
pathways; we conclude that the evolution of smRNA pathways has been a dynamic process that could generate
signatures of their diversification of function in plants. We have extended our investigations to the wheat genomes to
identify, annotate and understand signaling pathways and to determine phyloenetic linkages in other grass genomes. Our
results have direct implications for biotechnological applications for crop improvement in cereals in general and wheat
in particular.

CONFERENCE ABSTRACT (IN PROCESS)

TOLERANCE OF GRAFTS

A.P. Malyshkin

Orenburg State Medical Academy, 460000 Orenburg, Russia

Abstract

Studies on computer simulation of the genome (see, e.g., [1]) lead to understanding that not only the
characteristics of body elements (structure, weight, color, etc.), but also all interactions between them
are directly determined at the genomic level. Tolerance of self-antigens should also be directly
determined by the genome, through genomic or "smart" recognition, rather than through negative
selection against self-reactive lymphocytes. The concept of linked functions [2] holds that the very
presence of the genes of class I MHC self-antigens in the genome "automatically" precludes immune response to these
antigens. Therefore, integration of certain class I MHC genes of the donor into the genome of the recipient's
hematopoietic stem cells in the course of preoperative treatment should result in tolerance of the donor's MHC antigens.
Conceivably, this approach to the formation of tolerance should also work for xenogenic grafts, which would
considerably enhance the possibilities of tissue and organ transplantation. The technique for integrating foreign class I
MHC genes into the genome of hematopoietic stem cells has not been developed thus far. Solution of the numerous
problems involved requires experimental research. This research is going to take much effort, and I would appreciate
any suggestions on collaboration and/or sponsorship.

OMEGA-3: METABOLIC ENGINEERING NEW FATTY ACIDS FOR THE OILSEED INDUSTRY

Abstract

Omega-3 long chain polyunsaturated fatty acids like EPA and DHA have critical roles in human
health and development with studies indicating that deficiencies in these fatty acids can increase the
risk or severity of cardiovascular and inflammatory diseases in particular. These fatty acids are
predominantly sourced from fish and algal oils. In order to meet the increasing demand for these oils
there is an urgent need for an alternative and sustainable source of EPA and DHA. We have focused on maximising the
production of DHA in seed. This talk will describe the transition of DHA production in seed of our model species
Arabidopsis [1] through to Camelina [2], Brassica juncea and our target crop Brassica napus. DHA levels that exceed
the amount typically found in bulk fish oil have now been achieved in all four species. This talk will describe gene
characterisation, construct designs, transgenic plants and seed oil fatty acid profiles. We will describe the progress of the
ongoing GM canola field trials conducted by Nuseed as well as the effects of growth temperature on the performance of
transgenic and endogenous desaturases. We will also describe some of the intellectual property challenges this project
faced and how we have solved them to enable the real-world deployment of this new, high value GM crop.

CONFERENCE ABSTRACT (IN PROCESS)

MORPHO-GENETIC SCREENING OF THE PROMISING RICE GENOTYPES UNDER
SALINITY STRESS

State Key Laboratory of Rice Biology, China National Rice Research Institute, China

Abstract

Selection of new rice (Oryza sativa L.) variety tolerant to soil salinity is one of the most important
issues to avoid salt stress in rice production. In this study, the morphological characterization was
performed on 5 rice genotypes under two salt stress treatments. Morphological characters and
Na+/K+ uptake ratio were surveyed to evaluate salt stress effects. Under EC 12 dS/m of salt stress at
seedling stage, NSIC Rc222 and IR64 were found sensitive to salt compared to BRRI dhan47, Binadhan-8, and
Binadhan-10, which were moderately tolerant. Yet, at vegetative stage all the genotypes have survived. However, at
reproductive stage all the genotypes were affected by salt stress except for the Binadhan-10 that survived the whole life
cycle. A total of 160 SSR markers used have revealed 209 alleles among the 5 rice genotype. Interestingly, 4 SSR
markers with highest PIC (0.67) value have scored the highest level of genetic diversity value (0.72). The present
investigation on variant salt stress responsiveness of 5 promising rice genotype will be of great value in rice breeding.

Abstract

The detection of low-abundance biomarkers and targeted therapy of cancers using fabricated hybrid
nano-bioprobe are particularly desirable but remain a great challenge. In current work, designing
signal amplifying electrochemical biosensors by silicon carrier nanoparticles integrated with hybrid
gold nanoparticle-graphene nanosheet matrix became one appropriate way to overcome the drawback
of universal methods such as time consuming and insensitivity. By this manner, the combination of electrochemical and
luminescent methods has been proved to be a powerful analytical technique for ultrasensitive immunosensor and early
diagnosis with a detection limit up to 0.40 pg mL-1.

On the other hand, fabricating drug carrier with excellent performance, another remarkable coating strategy for I-III-VI
semiconductor quantum dots, the most promising and challenging QDs, has been achieved. In this regard, we
incorporated multiple hydrophobic CuInS2/ZnS and CdSe/CdS/ZnS directly into silica beads and further TEOS coating
via Stöber method was carried out. Afterwards, a mesoporous nanostructure was established via CTAB/NaOH mixture
with an overall size 45-50 nm in diameter, achieving a porous size 4 nm. More interesting, the addressed mesoporous
QDs turned to be a promising drug delivery carrier in the biomedical application and cancer therapy.

RESONANCE RAYLEIGH SCATTERING, SECOND-ORDER SCATTERING AND FREQUENCY
DOUBLING SCATTERING SPECTRA OF COPPER(II)-FLUTAMIDE SYSTEM WITH ANIONIC
SURFACTANTS AND ITS ANALYTICAL APPLICATION

Abstract

A simple, sensitive, and rapid method based on ion association, for the determination of FLD has
been developed. Flutamide (FLD) can react with Cu(II) to form 1:1 cationic chelate at pH 2.2-7.0
Mclivaine buffer medium, which can further react with anionic surfactants (AS) such as sodium
dodecyl sulfate (SDS), sodium lauryl sulfonate (SLS) and sodium dodecylbenzene sulfonate (SDBS)
to form 1:1 ion association complexes. As a result, the resonance Rayleigh scattering (RRS), second-order scattering
(SOS) and frequency doubling scattering (FDS) were enhanced to the highest degree. The maximum RRS, SOS and
FDS wavelengths of three ion-association complexes were located at 345/345 nm, 610/305 nm and 430/ 860 nm,
respectively. The increments of scattering intensity (ΔI) were directly proportional to the concentration of FLD in certain
ranges. The detection limits (3σ) of FLD for SDS, SLS and SDBS systems were 1.9 ng ml−1, 2.1 ng ml−1 and 2.2 ng
ml−1(RRS method), 2.4 ng ml−1, 2.7 ng ml−1 and 2.6 ng ml−1 (SOS method) and 2.3 ng ml−1, 2.4 ng ml−1 and 2.5 ng
ml−1 (FDS method), separately. The sensitivity of RRS method was higher than those of FDS and SOS methods. The
optimum conditions of RRS method and the influence factors, the composition and the reaction mechanism have been
discussed. Since the method is highly selective, it does not interference concomitant substances. These methods were
applied successfully for the determination of FLD in pharmaceutical formulation and urine.